Advances of Stem Cell therapy in Bone Regenration, Sinus Augmentation and others.

1. SMILE CONNECT REVIEW
AN UPDATE ON ADVANCES
IN STEM CELL THERAPY

2. 02
Index
Dental Stem Cells Compared to Bone Marrow . . . . . . . .
Dental Stem Cells as a source . . . . . . . .
Dental Stem Cells for tooth /Organ regeneration . . . . . . . .
Dental Stem Cells for Bone regeneration . . . . . . . .
Stem Cells used for Sinus Augmentation . . . . . . . .
Stem Cells for Orthopaedic Applications . . . . . . . .
Stem Cells in Opthalmic Applications . . . . . . . .
Stem Cells in Cardiac Applications . . . . . . . .
Stem Cells used for Diabetes Treatment . . . . . . . .
Neurological applications of Stem Cells . . . . . . . .
Immuno-modulatory properties of Stem Cells used in Therapy . . . . . . . .
Stem cells used to treat Liver conditions . . . . . . . .
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08
10
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3. 03
DentalStemCellsproventobeabettersourceof
Mesenchymalstemcellsthanbonemarrow
Mesenchymal Stem Cells Derived from Dental Tissues vs. Those from Other
Sources: Their Biology and Role in Regenerative Medicine
CRITICAL REVIEWS IN ORAL Biology & Medicine
G.T.-J. Huang1*, S. Gronthos2, and S. Shi3
Abstract
To date, 5 different human dental stem/progenitor cells have been isolated and characterized: dental pulp
stem cells (DPSCs), stem cells from exfoliated deciduous teeth (SHED), periodontal ligament stem cells
(PDLSCs), stem cells from apical papilla (SCAP), and dental follicle progenitor cells (DFPCs). These post-
natal populations have mesenchymal- stem-cell-like (MSC) qualities, including the capacity for self-renewal
and multilineage differentiation potential. MSCs derived from bone marrow (BMMSCs) are capable of giving
rise to various lineages of cells, such as osteogenic, chondrogenic,adipogenic, myogenic, and neurogenic
cells. The dental-tissue-derived stem cells are isolated from specialized tissue with potent capacitiesto
differentiate into odontogenic cells. However,they also have the ability to give rise to other cell lineages similar
to, but different in potency from,that of BMMSCs. This article will review the isolation and characterization of
the properties of different dental MSC-like populations in comparison with those of other MSCs, such as
BMMSCs. Important issues in stem cell biology, such as stem cell niche, homing, and immunoregulation, will
also be discussed.
1University of Maryland, College of Dental Surgery, Dental School, Department of Endodontics, Prosthodontics and Operative
Dentistry, 650 West Baltimore St., Baltimore, MD 21201, USA;
2Mesenchymal Stem Cell Group, Division of Haematology, Institute of Medical and Veterinary Science, Adelaide 5000, South
Australia, Australia; and 3Center for Craniofacial Molecular Biology, University of Southern California School of Dentistry, Los
Angeles, CA90033, USA; *corresponding author,ghuang@umaryland.edu J Dent Res 88(9):792-806, 2009
1
SHED: Stem cells from human exfoliated deciduous teeth
To isolate high-quality human postnatal stem cells from accessible resources is an important goal for stem-
cell research. In this study we found that exfoliated human deciduous tooth contains multi-potent stem cells
[stem cells from human exfoliated deciduous teeth (SHED)]. SHED were identified to be a population of
highlyproliferative, clonogenic cells capable of differentiating into a variety of cell types including neural cells,
adipocytes, and odon-toblasts. After in vivo transplantation, SHED were found to be able to induce bone
formation, generate dentin, and survive in mouse brain along with expression of neural markers. Here we
show that a naturally exfoliated human organ contains a population of stemcells that are completely different
from previously identified stem cells. SHED are not only derived from a very accessible tissue resource but
are also capable of providing enough cells for poten- tial clinical application. Thus, exfoliated teeth may be an
unexpected unique resource for stem-cell therapies including autologous stem-cell transplantation and
tissue engineering.
www.pnas.orgycgiydoiy10.1073ypnas.0937635100 PNAS u May 13, 2003 u vol. 100 u no. 10 u 5807–5812
Masako Miura*, Stan Gronthos†, Mingrui Zhao‡, Bai Lu‡, Larry W. Fisher*, Pamela Gehron Robey*, and Songtao Shi*§
*Craniofacial and Skeletal Diseases Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health,
Bethesda, MD 20892; †Mesenchymal Stem Cell Group, Division of Haematology, Institute of Medical and Veterinary Science, Frome
Road, Adelaide 5000, South Australia, Australia; and ‡Section on Neural Development and Plasticity, National Institute of Child Health
and Human Development, National Institutes of Health, Bethesda, MD 20892 Edited by Anthony P. Mahowald, University of Chicago,
Chicago, IL, and approved March 12, 2003 (received for review December 16, 2002)

4. Stem cell (SC) therapy has a promising future for tissue regenerative medicine. However, because SC technology
is still in its infancy, interdisciplinary cooperation is needed to achieve successful clinical applications. Dental SCs
have drawn attention in recent years because of their accessibility, plasticity, and high proliferative ability. Several
types of dental SCs have been identified, including dental pulp SCs from adult human dental pulp, SCs from
human primary exfoliated deciduous teeth, periodontal ligament SCs, and dental follicle SCs from human third
molars. Similar to mesenchymal SCs, these dental SCs can undergo self-renewal and have multipotent
differentiation ability, but do not have the ethical issues associated with other sources of SCs. Therefore,
appropriate preservation procedures for dental SCs and teeth are now needed. Here, we discuss the
opportunities for tooth-banking (as it is now clinically feasible and commercially available), the advantages and
limitations of current cryopreservation techniques for dental SCs/teeth or tissues, and the current status of tooth
banks.
Dental Stem Cells and Tooth Banking for Regenerative Medicine
J Exp Clin Med 2010;2(3):111–117
Yen-Hua Huang1,5, Jen-Chang Yang2,5, Chin-Wei Wang3,4, Sheng-Yang Lee3,4,5*1Department of Biochemistry, Graduate Institute of
Medical Sciences, School of Medicine; Center for Reproductive Medicine, Taipei Medical University Hospital, Taipei Medical University,
Taipei, Taiwan 2School of Dental Technology, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan 3Department of
Dentistry, Wan-Fang Hospital, Taipei Medical University, Taipei, Taiwan 4School of Dentistry, Taipei Medical University, Taipei, Taiwan
5Center for Teeth Bank and Dental Stem Cell Technology, Taipei Medical University, Taipei, Taiwan
DecidousTeethpromisingandimportantsourcefor
regenerativetherapy
Cryopreserved dental pulp tissues of exfoliated deciduous teeth is a feasible
stemcell resource for regenerative medicine.
Human exfoliated deciduous teeth have been considered to be a promising source for regenerative therapy
because they contain unique postnatal stem cells from human exfoliated deciduous teeth (SHED) with self-
renewal capacity, multipotency and immunomodulatory function. However preservation technique of deciduous
teeth has not been developed. This study aimed to evaluate that cryopreserved dental pulp tissues of
humanexfoliated deciduous teeth is a retrievable and practical SHED source for cell-based therapy. SHED
isolated from the cryopreserved deciduous pulp tissues for over 2 years (25-30 months) (SHED-Cryo) owned
similarstem cell properties including clonogenicity, self-renew, stem cell marker expression, multipotency, in vivo
tissue regenerative capacity and in vitro immunomodulatory function to SHED isolated from the fresh tissues
(SHED-Fresh). To examine the therapeutic efficacy of SHED-Cryo on immune diseases, SHED-Cryo were
intravenously transplanted into systemic lupus erythematosus (SLE) model MRL/lpr mice. Systemic SHED-Cryo-
transplantation improved SLE-like disorders including short lifespan, elevated autoantibody levels and nephritis-
like renal dysfunction. SHED-Cryo amended increased interleukin 17-secreting helper T cells in MRL/lpr mice
systemically and locally. SHED-Cryo-transplantation was also able to recover osteoporosis bone reduction in
long bones of MRL/lpr mice. Furthermore, SHED-Cryo-mediated tissue engineering induced bone regeneration
in critical calvarial bone-defect sites of immunocompromised mice. The therapeutic efficacy ofSHED-Cryo
transplantation on immune and skeletal disorders was similar to that of SHED-Fresh. These data suggest that
cryopreservation of dental pulp tissues of deciduous teeth provide a suitable and desirable approach for stem
cell-based immune therapy and tissue engineering in regenerative medicine.
PLoS One. 2012;7(12):e51777. doi: 10.1371/journal.pone.0051777. Epub 2012 Dec 14
Ma L, MakinoY,Yamaza H,Akiyama K, HoshinoY, Song G, KukitaT, Nonaka K, Shi S,YamazaT.
Department of Molecular Cell Biology and Oral Anatomy, Graduate School of Dental Science, Kyushu University, Fukuoka, Japan ;
Department of Pediatric Dentistry, Graduate School of Dental Science, Kyushu University, Fukuoka, Japan.
4

5. Summary： Human adult mesenchymal stem cells (MSCs)are rare elements living in various organs
(e.g.bone marrow,skeletal muscle),with capability to differentiate in various cell types (e.g.
chondrocytes,adipocytes and osteoblasts).In the year 2000,Gronthos and co-workers isolated stem
cells from the human dental pulp (DPSCs).Later on,stem cells from exfoliated tooth were also obtained.
The aims of our study were to establish protocol of DPSCs isolation and to cultivate DPSCs either from
adult or exfoliated tooth,and to compare these cells with mesenchymal progenitor cell (MPCs)cultures.
MPCs were isolated from the human bone marrow of proximal femur.DPSCs were isolated from
deciduous and permanent teeth.Both cell types were cultivated under the same conditions in the media
with 2 % of FCS supplemented with PDGF and EGF growth factors.We have cultivated
undifferentiated DPSCs for long time,over 60population doublings in cultivation media designed for
bone marrow MPCs.After reaching Hayflick’s limit,they still have normal karyotype.Initial doubling time
of our cultures was from 12to 50hours for first 40population doublings,after reaching 50population
doublings,doubling time had increased to 60– 90hours.Regression analysis of uncumulated
population doublings proved tight dependence of population doublings on passage number and slow
decrease of proliferation potential.In comparison with bone marrow MPCs,DPSCs share similar
biological characteristics and stem cell properties.The results of our experiments proved that the
DPSCs and MPCs are highly proliferative,clonogenic cells that can be expanded beyond Hayflick’s
limit and remain cytogenetically stable.Moreover we have probably isolated two different populations
of DPSCs.These DPSCs lines differed one from another in morphology.Because of their high
proliferative and differentiation potential,DPSCs can become more attractive,easily accessible source
of adult stem cells for therapeutic purposes.
Human Dental Pulp Stem Cells – Isolation and Long Term
Cultivation
` 1 ~ 2 1 2 1 3
Jakub Such anek ,Toma`s ,Romana Ivanèakov ,Jana Karbanov ,VìraHubkov ,Robert Pytlik ,
` 4
Lenka Kuèerov a
Charles University in Prague,Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove,Czech Republic,
Department of Dentistry1,Department of Histology and Embryology2,Department of Clinical Genetics4;Charles
University in Prague, 1st Medical Faculty in Prague,Teaching Hospital,Czech Republic: 1st Department of Medicine3
` ` `
a a a
DentalPulppresentsaneasyaccessible
sourceofstemcells
Abstract
Dental pulp (DP) can be extracted from child's primary teeth (deciduous), whose loss occurs spontaneously by
about 5 to 12 years. Thus, DP presents an easy accessible source of stem cells without ethical concerns.
Substantial quantities of stem cells of an excellent quality and at early (2-5) passages are necessary for clinical
use, which currently is a problem for use of adult stem cells. Herein, DPs were cultured generating stem cells at
least during six months through multiple mechanical transfers into a new culture dish every 3-4 days. We
compared stem cells isolated from the same DP before (early population, EP) and six months after several
mechanical transfers (late population, LP). No changes, in both EP and LP, were observed in morphology,
expression of stem cells markers (nestin, vimentin, fibronectin, SH2, SH3 and Oct3/4), chondrogenic and
myogenic differentiation potential, even after cryopreservation. Six hours after DP extraction and in vitro plating,
rare 5-bromo-2'-deoxyuridine (BrdU) positive cells were observed in pulp central part. After 72 hours, BrdU
positive cells increased in number and were found in DP periphery, thus originating a multicellular population of
stem cells of high purity. Multiple stem cell niches were identified in different zones of DP, because abundant
expression of nestin, vimentin and Oct3/4 proteins was observed, while STRO-1 protein localization was
restricted to perivascular niche. Our finding is of importance for the future of stem cell therapies, providing scaling-
up of stem cells at early passages with minimum risk of losing their "stemness".
PMCID: PMC3387222
Scaling-up of dental pulp stem cells isolated from multiple niches.
PLoS One. 2012;7(6):e39885. doi: 10.1371/journal.pone.0039885. Epub 2012 Jun 29.
Lizier NF, KerkisA, Gomes CM, Hebling J, Oliveira CF, CaplanAI, Kerkis I.
Laboratory of Genetics, Butantan Institute, Sao Paulo, SP, Brazil. nelson_lizier@yahoo.com.br
Stemade Comments: Dental Stem Cells have shown excellent potential for
expansion and culturing without loosing their “Stemness”
5

6. Abstract
In this study we used a biocomplex constructed from dental pulp stem/progenitor cells (DPCs) and a collagen
sponge scaffold for oro-maxillo-facial (OMF) bone tissue repair in patients requiring extraction of their third
molars. The experiments were carried out according to our Internal Ethical Committee Guidelines and written
informed consent was obtained from the patients. The patients presented with bilateral bone reabsorption of
the alveolar ridge distal to the second molar secondary to impaction of the third molar on the cortical alveolar
lamina, producing a defect without walls, of at least 1.5 cm in height. This clinical condition does not permit
spontaneous bone repair after extraction of the third molar, and eventually leads to loss also of the adjacent
second molar. Maxillary third molars were extracted first for DPC isolation and expansion. The cells were then
seeded onto a collagen sponge scaffold and the obtained biocomplex was used to fill in the injury site left by
extraction of the mandibular third molars. Three months after autologous DPC grafting, alveolar bone of
patients had optimal vertical repair and complete restoration of periodontal tissue back to the second molars,
as assessed by clinical probing and X-rays. Histological observations clearly demonstrated the complete
regeneration of bone at the injury site. Optimal bone regeneration was evident one year after grafting. This
clinical study demonstrates that a DPC/collagen sponge biocomplex can completely restore human
mandible bone defects and indicates that this cell population could be used for the repair and/or regeneration
of tissues and organs.
Human Mandible Bone Defect Repair by The Grafting of Dental Pulp Stem/
progenitor Cells and Collagen Sponge Biocomplexes
Riccardo d’Aquino1,2, Alfredo De Rosa1, Vladimiro Lanza1, Virginia Tirino2, Luigi Laino1, Antonio Graziano1, Vincenzo
Desiderio2, Gregorio Laino1 and Gianpaolo Papaccio2*
Dipartimento di Discipline Odontostomatologiche, Ortodontiche e Chirurgiche, 2Dipartimento di Medicina Sperimentale, Sezione di
Istologia ed Embriologia,Tissue Engineering and Regenerative Medicine (TERM) Division, SecondoAteneo di Napoli, Naples, Italy
9
DentalStemCellsforRepairofMandible
BoneDefect
DentalStemCellsuseforBoneRegeneration
Stromal stem cells from human dental pulp (SBP-DPSCs) were used to study osteogenic differentiation in
vitro and in vivo. We previously reported that SBP-DPSCs are multipotent stem cells able to differentiate into
osteoblasts, which synthesize three dimensional woven bone tissue chips in vitro. In this study, we followed
the temporal expression pattern of speci?c markers in SBP-DPSCs and found that, when differentiating into
osteoblasts, they express, besides osteocalcin, also ?k-1 (VEGF-R2). In addition, 30% of them expressed
speci?c antigens for endothelial cells, including CD54, von-Willebrand (domain 1 and 2), CD31 (PECAM-1)
and angiotensin-converting enzyme. Interestingly, we found endotheliocytes forming vessel walls, observing
that stem cells synergically differentiate into osteoblasts and endotheliocytes, and that ?k-1 exerts a pivotal
role in coupling osteoblast and endotheliocyte differentiation. When either SBP-DPSCs or bone chips
obtained in vitro were transplanted into immunocompromised rats, they generated a tissue structure with an
integral blood supply similar to that of human adult bone; in fact, a large number of HLA-1 þ vessels were
observed either within the bone or surrounding it in a periosteal layer. This study provides direct evidence to
suggest that osteogenesis and angiogenesis mediated by human SBP-DPSCs may be regulated by distinct
mechanisms, leading to the organization of adult bone tissue after stem cell transplantion.
Cell Death and Differentiation (2007) 14, 1162–1171. doi:10.1038/sj.cdd.4402121; published online 9 March 2007
Human postnatal dental pulp cells co-differentiate into osteoblasts and
endotheliocytes: a pivotal synergy leading to adult bone tissue formation
R d’Aquino1,2,5,AGraziano2,5, M Sampaolesi3, G Laino1, G Pirozzi4,ADe Rosa1 and G Papaccio*,2
8

7. Sinus Lift Augmentation Using Autologous Pulp Stem Cells:
Case Report Of Bone Density Evaluation
Bone augmentation to reconstruct atrophic jaws provides the base for sufficient functional and aesthetic
implant-supported oral rehabilitation. Although autografts are the standard procedure for bone grafting, the
use of bone regeneration by means of dental pulp stem cell is an alternative that open a new era in this field. In
March 2010, at the Department of Oral Surgery, Don Orione Hospital, Bergamo, Italy, one patient undergo to
sinus lift elevation with pulp stem cells gentle poured onto collagen sponge. A CT scan control was
performed after 4 month and DICOM data were processed with medical imaging software which gives the
possibility to use a virtual probe to extract the bone density. Pearson’s chi-square test was used to investigate
difference in bone density (i.e. BD) between native and newly formed bone. BD in newly formed bone is about
the double of native bone. This report demonstrated that stem cells derived from dental pulp poured onto
collagen sponge is a useful method for bone regeneration in atrophic maxilla.
G. BRUNELLI1,A. MOTRONI2, F. CARINCI3,A. GRAZIANO3,
R. D’AQUINO3, I. ZOLLINO3, R. MONGUZZI3,
Don Orione Hospital, Bergamo, Italy
2AMIRG (Applied Medical Imaging Research Group), Milan, Italy
3 Department of D.M.C.C.C., Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy
Lancet. 2008 May 10;371(9624):1579-86. doi: 10.1016/S0140-6736(08)60690-X.
11
Pleasemailthebulletpoint
MesenchymalStemCellsusedforSinus
augmentation
PURPOSE: Mesenchymal stem cells (MSCs) have been applied in maxillary sinus augmentation (MSA) with
clinically successful results. The purpose of this article was to evaluate the systematically acquired evidence for
the effectiveness of cell-based approaches in MSA with various scaffolds, and to narratively assess evidence
from additional articles that report effectiveness of cell-based approaches in MSA.
MATERIALS AND METHODS: Electronic database searches were performed. Inclusion criteria were studies
of cell-based approaches in MSA with various scaffolds, in humans, with at least 3 to 4 months of follow-up.
Meta-analysis was performed for randomized controlled trials (RCTs) with histologic/histomorphometric
evaluation.
RESULTS: Fifteen studies (4 RCTs) were considered to be eligible for inclusion in the review. The meta-analysis
suggested a marginal, nonstatistically significant positive effect of MSCs on the bone regrowth.
CONCLUSIONS: A number of studies have demonstrated the potential for cell-based approaches in MSA;
further RCTs that clearly demonstrate benefits of cell-based approach are needed.
Maxillary sinus augmentation with adult mesenchymal stem cells: a review of the
current literature.
Oral Surg Oral Med Oral Pathol Oral Radiol. 2013 Jan 9. pii: S2212-4403(12)01627-6. doi: 10.1016/j.oooo.2012.09.087. Mangano
FG,Tettamanti L, Sammons RL,Azzi L, CaprioglioA, MacchiA, Mangano C
Dental School, University of Insubria, Varese, Italy. Electronic address: francescomangano1@mclink.net.
10

8. Abstract
The need for new dental tissue-replacement therapies is evident in recent reports which reveal startling
statistics regarding the high incidence of tooth decay and tooth loss. Recent advances in the identification
and characterization of dental stem cells, and in dental tissue-engineering strategies, suggest that
bioengineering approaches may successfully be used to regenerate dental tissues and whole teeth. Interest
in dental tissue-regeneration applications continues to increase as clinically relevant methods for the
generation of bioengineered dental tissues, and whole teeth, continue to improve. This paper is concerned
about dental-derived stem cells and their characterization. Additionally, since conventional dental treatments
partially serve the purpose for replacing missing teeth and always include possible failure rates, the potential
of dental-derived stem cells in promoting whole tooth regeneration is also discussed.
Dental-derived Stem Cells and whole Tooth Regeneration: an Overview
Aous Dannan
DentalStemCellsforwhole
ToothRegeneration
6
Repair of alveolar cleft defect with mesenchymal stem cells and platelet
derived growth factors: a preliminary report.
The purpose of this study was to evaluate the enhancing effect of recombinant platelet derived growth factor
on human mesenchymal stem cells(hMSCs) in secondary alveoloplasty. Three patients with 4 alveolar
defects were selected for this study. Mesenchymal stem cells were cultured from a posterior iliac bone
aspirate. MSCs were mounted on biphasic scaffolds and combined with platelet derived growth factor
(PDGF) in the operating room to make a triad of the scaffold, growth factor, and cells. The triads were placed
in anterior maxillary cleft defects and closed with lateral advancement gingival flaps. The postoperative cleft
bone volume was measured with cone beam computed tomography scans. A mean of 51.3% fill of the bone
defect was calculated 3 months post-operation. Our data suggests the use of recombinant platelet derived
growth factor with hMSCs may enhance the regeneration capacity of the cells.
J Craniomaxillofac Surg. 2012 Jan;40(1):2-7. doi: 10.1016/j.jcms.2011.02.003. Epub 2011 Mar 21. Behnia H, Khojasteh A,
Soleimani M,TehranchiA,AtashiA.
Department of Oral and Maxillofacial Surgery, Dental Research Center, Shahid Beheshti University of Medical Sciences, Tehran,
Iran.
MesenchymalStemCellsusedto
treatAlveolarcleftdefect
12

9. PMID: 19614680 [PubMed - indexed for MEDLINE]
DentalStemCellshavingsamecharacteristicsaslimbalstemcells
Human immature dental pulp stem cells share key characteristic features with
limbal stem cells.
ABSTRACT
OBJECTIVES: Limbal stem cells (LSC) are self-renewing, highly proliferative cells in vitro, which express a set of
specific markers and in vivo have the capacity to reconstruct the entire corneal epithelium in cases of ocular
surface injury. Currently, LSC transplantation is a commonly used procedure in patients with either uni- or bilateral
total limbal stem cells deficiency (TLSCD). Although LSC transplantation holds great promise for patients, several
problems need to be overcome. In order to find an alternative source of cells that can partially substitute LSC in
cornea epithelium reconstruction, we aimed at investigating whether human immature dental pulp stem cells
(hIDPSC) would present similar key characteristics as LSC and whether they could be used for corneal surface
reconstruction in a rabbit TLSCD model.
MATERIALS: We used hIDPSC, which co-express mesenchymal and embryonic stem cell markers and present
the capacity to differentiate into derivative cells of the three germinal layers. TLSCD was induced by chemical burn
in one eye of rabbits. After 30 days, the opaque tissue formed was removed by superficial keratectomy.
Experimental group received undifferentiated hIDPSC, while control group only received amniotic membrane
(AM). Both groups were sacrificed after 3 months.
RESULTS AND CONCLUSIONS: We have demonstrated, using immunohistochemistry and reverse
transcription-polymerase chain reaction, that hIDPSCs express markers in common with LSC, such as ABCG2,
integrin beta1, vimentin, p63, connexin 43 and cytokeratins 3/12. They were also capable of reconstructing the
eye surface after induction of unilateral TLSCD in rabbits, as shown by morphological and immunohistochemical
analysis using human-specific antibodies against limbal and corneal epithelium. Our data suggest that hIDPSCs
share similar characteristics with LSC and might be used as a potential alternative source of cells for corneal
reconstruction.
Cell Prolif. 2009 Oct;42(5):587-94. Epub 2009 Jul 14.
Monteiro BG, Serafim RC, Melo GB, Silva MC, Lizier NF, Maranduba CM, Smith RL, KerkisA, Cerruti H, Gomes JA, Kerkis I.
Source: Genetics Laboratory, Butantan Institute, São Paulo, SP, Brazil.
20
DentalPulpMesenchymalStemCellsreconstructcornea
Corneal Reconstruction with Tissue-Engineered Cell Sheets Composed of
Human Immature Dental Pulp Stem Cells
PURPOSE: To determine the outcome of the use of a tissue-engineered cell sheet composed of human
undifferentiated immature dental pulp stem cells (hIDPSC) for ocular surface reconstruction in an animal model of
total limbal stem cell deficiency (LSCD).
METHODS: LSCD was induced by the application of 0.5 M NaOH to the right eye of rabbits for 25 seconds (mild
chemical burn [MCB]) and for 45 seconds (severe chemical burn [SCB]). After 1 month, a superficial keratectomy
was performed to remove the fibrovascular pannus that covered the animals' burned corneas. A tissue-
engineered hIDPSC sheet was transplanted onto the corneal bed and then covered with deepithelialized human
amniotic membrane (AM). In the respective control groups, the denuded cornea was covered with AM only. After 3
months, a detailed analysis of the rabbit eyes was performed with regard to clinical aspect, histology, electron
microscopy, and immunohistochemistry.
RESULTS: Corneal transparency of the rabbit eyes that underwent hIDPSC transplantation was improved
throughout the follow-up, while the control corneas developed total conjunctivalization and opacification. Rabbits
from the MCB group showed clearer corneas with less neovascularization. The clinical data were confirmed by
histologic analysis that showed healthy uniform corneal epithelium, especially in the MCB group. The presence of
hIDPSC was detected using an anti- hIDPSC antibody. The corneal tissue also showed positive
Investigative Ophthalmology & Visual Science, March 2010, Vol. 51, No. 3
Jose´ A´ lvaro Pereira Gomes,*,1 Ba´byla Geraldes Monteiro,2,3 Gustavo Barreto Melo,1 Ricardo Luiz Smith,4 Marcelo Cavenaghi
Pereira da Silva,4 Nelson Foresto Lizier,3,4Alexandre Kerkis,5 Humberto Cerruti,6 and Irina Kerkis*,3
1Centro Avanc¸ado de Superfície Ocular (CASO) and Setor de Co´rnea e Doenc¸as Externas, Instituto da Visa˜o, Universidade Federal
de Sa˜o Paulo, Sa˜o Paulo, Brazil; 3Laborato´rio de Gene´tica, Instituto Butan-tan, Instituto Nacional de Cieˆncia e Tecnologia em
Ce´lulas-Tronco eTerapia Celular (INCTC), Sa˜o Paulo, Brazil; 4Departamento de Morfo-logia e Gene´tica da Universidade Federal de
Sa˜o Paulo, Sa˜o Paulo,Brazil; 5Gene´tica Aplicada, Atividades Veterina´rias Ltda., Centro deInovac¸a˜o Tecnolo´gica, Sa˜o Paulo,
Brazil; and 6Clínica e Centro de Pesquisa Odontolo´gica-CERA, Sa˜o Paulo, Brazil.
21

10. AutologousOralMucosalEpitheliumforCorneal
Reconstruction
BACKGROUND: Ocular trauma or disease may lead to severe corneal opacification and, consequently, severe
loss of vision as a result of complete loss of corneal epithelial stem cells. Trans-plantation of autologous corneal
stem-cell sources is an alternative to allograft trans-plantation and does not require immunosuppression, but it is
not possible in many cases in which bilateral disease produces total corneal stem-cell deficiency in both eyes. We
studied the use of autologous oral mucosal epithelial cells as a source of cells for the reconstruction of the corneal
surface.
METHODS: We harvested 3-by-3-mm specimens of oral mucosal tissue from four patients with bilateral total
corneal stem-cell deficiencies. Tissue-engineered epithelial-cell sheets were fabricated ex vivo by culturing
harvested cells for two weeks on temperature-responsive cell-culture surfaces with 3T3 feeder cells that had been
treated with mi-tomycin C. After conjunctival fibrovascular tissue had been surgically removed from the ocular
surface, sheets of cultured autologous cells that had been harvested with a sim-ple reduced-temperature
treatment were transplanted directly to the denuded corneal surfaces (one eye of each patient) without sutures.
RESULTS: Complete reepithelialization of the corneal surfaces occurred within one week in all four treated eyes.
Corneal transparency was restored and postoperative visual acuity im-proved remarkably in all four eyes. During
a mean follow-up period of 14 months, all corneal surfaces remained transparent. There were no complications.
CONCLUSIONS: Sutureless transplantation of carrier-free cell sheets composed of autologous oral mu-cosal
epithelial cells may be used to reconstruct corneal surfaces and can restore vision in patients with bilateral severe
disorders of the ocular surface.
Corneal Reconstruction with Tissue- Engineered Cell Sheets Composed of
Autologous Oral Mucosal Epithelium
Kohji Nishida, M.D., Ph.D., Masayuki Yamato, Ph.D., Yasutaka Hayashida, M.D., Katsuhiko Watanabe, M.Sc., Kazuaki Yamamoto,
M.Sc., Eijiro Adachi, M.D., Ph.D., Shigeru Nagai, M.Sc., Akihiko Kikuchi, Ph.D., Naoyuki Maeda, M.D., Ph.D., Hitoshi Watanabe, M.D.,
Ph.D.,Teruo Okano, Ph.D., andYasuoTano, M.D., Ph.D.
n engl j med 351;12 www.nejm.org september 16, 2004
19
Mesenchymalcellsforcornealrecunstruction
ABSTRACT
PURPOSE: Ocular burns can damage the corneal epithelial stem cells located at the limbus. This study
evaluated the efficacy of xeno-free autologous cell-based treatment of limbal stem cell deficiency.
METHODS: This retrospective study included 200 patients, above 8 years of age, with clinically diagnosed
unilateral total limbal stem cell deficiency due to ocular surface burns treated between 2001 and 2010. A small
limbal biopsy was obtained from the unaffected eye. The limbal epithelial cells were expanded ex vivo on
human amniotic membrane for 10e14 days using a xeno-free explant culture system. The resulting cultured
epithelial monolayer and amniotic membrane substrate were transplanted on to the patient's affected eye.
Postoperative corneal surface stability, visual improvement and complications were objectively analysed.
RESULTS: A completely epithelised, avascular and clinically stable corneal surface was seen in 142 of 200
(71%) eyes at a mean follow-up of 361.6 (range: 1e7.6) years. A two-line improvement in visual acuity, without
further surgical intervention, was seen in 60.5% of eyes. All donor eyes remained healthy.
CONCLUSIONS: Autologous cultivated limbal epithelial transplantation using a xeno-free explant culture
Clinical outcomes of xeno-free autologous cultivated limbal epithelial
transplantation: a 10-year study
BJO 10.1136/bjophthalmol-2011-300352 Virender S Sangwan,1,2 Sayan Basu,1,2 Geeta K Vemuganti,2 Kunjal Sejpal,1 Sandhya
V Subramaniam,3 Souvik Bandyopadhyay,4 Sannapaneni Krishnaiah,5 Subhash Gaddipati,2,6 Shubha Tiwari,2,6 Dorairajan
Balasubramanian2,3,6
1Cornea and Anterior Segment Services, L V Prasad Eye Institute, Hyderabad, India 2Sudhakar and Sreekanth Ravi Stem Cell
Biology Laboratory, L V Prasad Eye Institute, Hyderabad, India 3Professor Brien Holden Eye Research Center, L V Prasad Eye
Institute, Hyderabad, India 4Indian Institute of Public Health, Hyderabad, India 5Department of Clinical Epidemiology and
Biostatistics, L V Prasad Eye Institute, Hyderabad, India 6Champalimaud Translational Center for Eye Research, LV Prasad Eye
Institute, Hyderabad, India
15

11. limbalepithelialtransplantationfortreatingunilateral
limbalstemcelldeficiencyfollowingocularburns.
Simple limbal epithelial transplantation (SLET): a novel surgical technique for
the treatment of unilateral limbal stem cell deficiency
ABSTRACT
This study describes a novel surgical technique of limbal transplantation, which combines the benefits of
existing techniques while avoiding their difficulties. Six patients with unilateral and total limbal stem cell
deficiency following ocular surface burns underwent a single-stage procedure. A 232 mm strip of donor limbal
tissue was obtained from the healthy eye and divided into eight to ten small pieces. After surgical preparation
of the recipient ocular surface, these tiny limbal transplants were distributed evenly over an amniotic
membrane placed on the cornea. After surgery, a completely epithelialised, avascular and stable corneal
surface was seen in all recipient eyes by 6 weeks, and this was maintained at a mean6SD follow-up of 9.261.9
months. Visual acuity improved from worse than 20/200 in all recipient eyes before surgery to 20/60 or better in
four (66.6%) eyes, while none of the donor eyes developed any complications. This technique requires less
donor tissue than previously used for conventional autografting and does not need a specialist laboratory for
cell expansion. Although long-term results are awaited, this simple limbal epithelial transplantation promises
to be an easy and effective technique for treating unilateral limbal stem cell deficiency following ocular burns.
Virender S Sangwan,1,2 Sayan Basu,1,2 Sheila MacNeil,3 Dorairajan Balasubramanian4 <Additional materials are published online
only. To view these files please visit the journal online (http://bjo.bmj. com/content/early/recent). 1Cornea and Anterior Segment
Services, L V Prasad Eye Institute, Hyderabad, India 2Sudhakar and Sreekanth Ravi Stem Cell Biology Laboratory, L V Prasad Eye
Institute, Hyderabad, India 3Department of Materials Science and Engineering, University of Sheffield, Sheffield, UK
4Champalimaud Translational Centre for Eye Research, Hyderabad Eye Research Foundation, L V Prasad Eye Institute,
Hyderabad, India
16
Intracoronary autologous bone-marrow cell transfer after myocardial infarction:
the BOOST randomised controlled clinical trial
SUMMARY
BACKGROUND: Emerging evidence suggests that stem cells and progenitor cells derived from bone marrow
can be used to improve cardiac function in patients after acute myocardial infarction. In this randomised trial, we
aimed to assess whether intracoronary transfer of autologous bone-marrow cells could improve global left-
ventricular ejection fraction (LVEF) at 6 months' follow-up.
METHODS: After successful percutaneous coronary intervention (PCI) for acute ST-segment elevation
myocardial infarction, 60 patients were randomly assigned to either a control group (n=30) that received
optimum postinfarction medical treatment, or a bone-marrow-cell group (n=30) that received optimum medical
treatment and intracoronary transfer of autologous bone-marrow cells 4·8 days (SD 1·3) after PCI. Primary
endpoint was global left-ventricular ejection fraction (LVEF) change from baseline to 6 months' follow-up, as
determined by cardiac MRI. Image analyses were done by two investigators blinded for treatment assignment.
Analysis was per protocol.
FINDINGS: Global LVEF at baseline (determined 3·5 days [SD 1·5] after PCI) was 51·3 (9·3%) in controls and 50·0
(10·0%) in the bone-marrow cell group (p=0·59). After 6 months, mean global LVEF had increased by 0·7
percentage points in the control group and 6·7 percentage points in the bone-marrow-cell group (p=0·0026).
Transfer of bone-marrow cells enhanced left-ventricular systolic function primarily in myocardial segments
adjacent to the infarcted area. Cell transfer did not increase the risk of adverse clinical events, in stent restenosis,
or proarrhythmic effects.
INTERPRETATION: Intracoronary transfer of autologous bone-marrow-cells promotes improvement of left-
ventricular systolic function in patients after acute myocardial infarction.
Kai C Wollert MD a, Gerd P Meyer MD a, Joachim Lotz MD b, Stefanie Ringes Lichtenberg MD a, Peter Lippolt MD a, Christiane
Breidenbach MD a, Stephanie Fichtner BS a, Thomas Korte MD a, Burkhard Hornig MD a, Diethelm Messinger MSc d, LubomirArseniev
MD c, Bernd Hertenstein MD c, ProfArnold Ganser MD c, Prof Helmut Drexler MD
The Lancet, Volume 364, Issue 9429, Pages 141 - 148, 10 July 2004
doi:10.1016/S0140-6736(04)16626-9
StemCellsimproveleft-ventricularsystolicfunction
afteracutemyocardialinfarction
22

12. 14
StemCellTherapyforMyocardialInfarctionStemCellTherapyimprovesventricularperformance,
qualityoflifeandsurvivalinpatientswithheartfailure
AIMS: We assessed whether a single intracoronary infusion of autologous bone marrow cells (BMCs) can have a
sustained impact on left ventricular ejection fraction (LVEF) in patients after ST-elevation myocardial infarction
(STEMI). In the BOne marrOw transfer to enhance ST-elevation infarct regeneration (BOOST) trial, 60 patients with
STEMI and successful percutaneous coronary intervention were randomized to a control and a cell therapy
group. As previously reported, BMC transfer led to an improvement of LVEF by 6.0% at 6 months (P ¼ 0.003) and
2.8% at 18 months (P ¼ 0.27).
METHODS: and results Left ventricular ejection fraction and clinical status were re-assessed in all surviving
patients after 61+11 months. Major adverse cardiac events occurred with similar frequency in both groups. When
compared with baseline, LVEF assessed by magnetic resonance imaging at 61 months decreased by 3.3+9.5%
in the control group and by 2.5+11.9% in the BMC group (P ¼ 0.30). Patients with an infarct transmurality . median
appeared to benefit from BMC transfer throughout the 61-month study period (P ¼ 0.040).
CONCLUSION: A single intracoronary application of BMCs does not promote a sustained improvement of LVEF
in STEMI patients with relatively preserved systolic function. It is conceivable that a subgroup of patients with
more transmural infarcts may derive a sustained benefit from BMC therapy. However, this needs to be tested
prospectively in a randomized trial.
KEYWORDS: Acute myocardial infarction • Cell therapy • Magnetic resonance imaging
Intracoronary bone marrow cell transfer after myocardial infarction: 5-year follow-
up from the randomized-controlled BOOST trial
Gerd P. Meyer1*†, Kai C. Wollert1†, Joachim Lotz2, Jens Pirr1, Ulrike Rager1, Peter Lippolt1, Andreas Hahn3, Stephanie
Fichtner1,Arnd Schaefer1, LubomirArseniev4,Arnold Ganser4, and Helmut Drexler1,
1Department of Cardiology andAngiology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany; 2Department of
Diagnostic Radiology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany; 3Department of Biometrics,
Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany; and 4Department of Hematology, Hemostaseology,
Oncology, and Stem Cell Transplantation, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany Received 15
October 2008; revised 19 July 2009; accepted 20August 2009; online publish-ahead-of-print 22 September 2009
AIMS: Despite accumulated evidence that intracoronary bone marrow cell (BMC) therapy may be beneficial in
acute myocardial infarction, there are only limited data available on the effectiveness of BMC’s in chronic heart
failure. The aim of this study was to quantitatively investigate ventricular haemodynamics, geometry, and
contractility as well as the long-term clinical outcome of BMC treated patients with reduced left ventricular ejection
fraction (LVEF) due to chronic ischaemic cardiomyopathy.
METHODS AND RESULTS: Patients with chronic heart failure (n ¼ 391 LVEF ≤ 35%) due to ischaemic
cardiomyopathy were enrolled in the present study. Of these, 191 patients (mean NYHA class 3.22) underwent
intracoronary BMC therapy. The control group (mean NYHA class 3.06) consisted of 200 patients with
comparable LVEF. Assessments of haemodynamics at rest and exercise, quantitative ventriculography,
spiroergometry, 24 h Holter ECG, late potentials, and heart rate variability were analysed. Over 3 months to 5
years after intracoronary BMC therapy there was a significant improvement in haemodynamics (e.g. LVEF,
cardiac index), exercise capacity, oxygen uptake, and LV contractility. Importantly, there was a significant
decrease in long-term mortality in the BMC treated patients compared with the control group.
CONCLUSION: Intracoronary BMC therapy improves ventricular performance, quality of life and survival in
patients with heart failure.These effects were present when BMC were administered in addition to standard
therapeutic regimes. No side effects were observed.
KEYWORDS: Ischaemic cardiomyopathy • Chronic heart failure • Bone marrow cell therapy • Remodelling •
Mortality
The acute and long-term effects of intracoronary Stem cell Transplantation in 191
patients with chronic heARt failure: the STAR-heart study
Bodo-Eckehard Strauer*, MuhammadYousef, and Christiana M. Schannwell
From the Department of Medicine, Division of Cardiology, Pneumology and Angiology, Heinrich-Heine-University of Du¨sseldorf,
Moorenstr 5, 40225 Du¨sseldorf, Germany Received 11 February 2010; revised 12April 2010; accepted 16April 2010
See page 642 for the editorial comment on this article (doi:10.1093/eurheartj/hfq105)
23
24

13. 15
Transplantation of Progenitor Cells and Regeneration Enhancement in Acute
Myocardial Infarction (TOPCARE-AMI)
ABSTRACT
BACKGROUND: Experimental studies suggest that transplantation of blood-derived or bone marrow–derived
progenitor cells beneficially affects postinfarction remodeling. The safety and feasibility of autologous progenitor
cell transplantation in patients with ischemic heart disease is unknown.
METHODS AND RESULTS: We randomly allocated 20 patients with reperfused acute myocardial infarction (AMI)
to receive intracoronary infusion of either bone marrow–derived (n=9) or circulating blood–derived progenitor
cells (n=11) into the infarct artery 4.3±1.5 days after AMI. Transplantation of progenitor cells was associated with
a significant increase in global left ventricular ejection fraction from 51.6±9.6% to 60.1±8.6% (P=0.003),
improved regional wall motion in the infarct zone (- 1.5±0.2 to - 0.5±0.7 SD/chord; P<0.001), and profoundly
reduced end-systolic left ventricular volumes (56.1±20 mL to 42.2±15.1 mL; P=0.01) at 4-month follow-up. In
contrast, in a nonrandomized matched reference group, left ventricular ejection fraction only slightly increased
from 51±10% to 53.5±7.9%, and end-systolic volumes remained unchanged. Echocardiography revealed a
profound enhancement of regional contractile function (wall motion score index 1.4±0.2 at baseline versus
1.19±0.2 at follow-up; P<0.001). At 4 months, coronary blood flow reserve was significantly (P<0.001) increased
in the infarct artery. Quantitative F-18-fluorodeoxyglucose–positron emission tomography analysis revealed a
significant (P<0.01) increase in myocardial viability in the infarct zone. There were no differences for any
measured parameter between blood-derived or bone marrow–derived progenitor cells. No signs of an
inflammatory response or malignant arrhythmias were observed.
CONCLUSIONS: In patients with AMI, intracoronary infusion of autologous progenitor cells appears to be
feasible and safe and may beneficially affect postinfarction remodeling processes.
Birgit Assmus, MD; Volker Schächinger, MD; Claudius Teupe, MD;Martina Britten, MD; Ralf Lehmann, MD; Natascha Döbert, MD;Frank
Grünwald, MD; Alexandra Aicher, MD; Carmen Urbich, PhD;Hans Martin, MD; Dieter Hoelzer, MD; Stefanie Dimmeler, PhD;Andreas M.
Zeiher, MD
Author Affiliations: From the Department of Molecular Cardiology and Department of Cardiology, Internal Medicine IV, (B.A., V.S., C.T.,
M.B., R.L., A.A., C.U.), Department of Nuclear Medicine (N.D., F.G.), and the Department of Hematology (H.M., D.H.), University of
Frankfurt, Frankfurt, Germany. Circulation.2002; 106: 3009-3017Published online before print November 11, 2002,doi:
10.1161/?01.CIR.0000043246.74879.CD
StemCellsUsedPostAcuteMyocardialInfarction
AIMS: To investigate the clinical outcome after intracoronary administration of autologous progenitor cells in
patients with acute myocardial infarction (AMI).
METHODS AND RESULTS: Using a double-blind, placebo-controlled multicentre trial design, we randomized
204 patients with successfully reperfused AMI to receive intracoronary infusion of bone-marrow-derived
progenitor cells (BMCs) or placebo medium into the infarct artery 3–7 days after successful infarct reperfusion
therapy. At 12 months, the pre-specified cumulative endpoint of death, myocardial infarction, or necessity for
revascularization was significantly reduced in the BMC group compared with placebo (P ¼ 0.009). Likewise, the
combined endpoint death, recurrence of myocardial infarction, and rehospitalization for heart failure was
significantly (P ¼ 0.006) reduced in patients receiving intracoronary BMC administration. Intracoronary
administration of BMC remained a significant predictor of a favourable clinical outcome by Cox regression
analysis, adjusting for classical predictors of poor outcome after AMI.
CONCLUSION: Intracoronary administration of BMCs is associated with a significant reduction of the occurrence
of major adverse cardiovascular events after AMI. Large-scale studies are warranted to confirm the effects of BMC
administration on mortality and morbidity in patients with AMIs.
KEYWORDS: Myocardial infarction; Prognosis; Cells;Catheterization
Improved clinical outcome after intracoronary administration of bone-marrow-
derived progenitor cells in acute myocardial infarction: final 1-year results of the
REPAIR-AMI trial
Volker Scha¨chinger1, Sandra Erbs2,Albrecht Elsa¨sser3, Werner Haberbosch4, Rainer Hambrecht2, Hans Ho¨lschermann5, Jiangtao
Yu6, Roberto Corti7, Detlef G. Mathey8, Christian W. Hamm3, Tim Su¨selbeck9, Nikos Werner10{, Ju¨rgen Haase11, Jo¨rg Neuzner12,
Alfried Germing13, Bernd Mark14, BirgitAssmus1, Torsten Tonn15, Stefanie Dimmeler16, andAndreas M. Zeiher1* for the REPAIR-AMI
Investigators
StemCellTherapyinTreatmentofAcute
MyocardialInfarction
25
26

14. Human dental pulp stem cells improve left ventricular function, induce
angiogenesis, and reduce infarct size in rats with acute myocardial infarction.
ABSTRACT
Human dental pulp contains precursor cells termed dental pulp stem cells (DPSC) that show self-renewal and
multilineage differentiation and also secrete multiple proangiogenic and antiapoptotic factors. To examine
whether these cells could have therapeutic potential in the repair of myocardial infarction (MI), DPSC were
infected with a retrovirus encoding the green fluorescent protein (GFP) and expanded ex vivo. Seven days
after induction of myocardial infarction by coronary artery ligation, 1.5 x 10(6) GFP-DPSC were injected
intramyocardially in nude rats. At 4 weeks, cell-treated animals showed an improvement in cardiac function,
observed by percentage changes in anterior wall thickening left ventricular fractional area change, in parallel
with a reduction in infarct size. No histologic evidence was seen of GFP+ endothelial cells, smooth muscle
cells, or cardiac muscle cellswithin the infarct. However, angiogenesis was increased relative to control-
treated animals. Taken together, these data suggest that DPSC could provide a novel alternative cell
population for cardiac repair, at least in the setting of acute MI.
Gandia C,ArmiñanA, García-Verdugo JM, Lledó E, RuizA, Miñana MD, Sanchez-Torrijos J, Payá R, Mirabet V, Carbonell-Uberos F,
Llop M, Montero JA,Sepúlveda P.
Source: Fundación Hospital General Universitario, Consorcio Hospital General Universitario de Valencia, Avenida Tres Cruces s/n,
46014 Valencia, Spain.
Stem Cells. 2008 Mar;26(3):638-45. Epub 2007 Dec 13.
DentalStemCellsimproveleftventricular
functionafteracutemyocardialinfarction
27
Stem cell therapy for type 1 diabetes mellitus: a review of recent
clinical trials
Abstract
Stem cell therapy is one of the most promising treatments for the near future. It is expected that this kind
of therapy can ameliorate or even reverse some diseases. With regard to type 1 diabetes, studies
analyzing the therapeutic effects of stem cells in humans began in 2003 in the Hospital das Clínicas of the
Faculty of Medicine of Ribeirão Preto - SP USP, Brazil, and since then other centers in different countries
started to randomize patients in their clinical trials. Herein we summarize recent data about beta cell
regeneration, different ways of immune intervention and what is being employed in type 1 diabetic
patients with regard to stem cell repertoire to promote regeneration and/or preservation of beta cell mass.
The Diabetes Control and Complications Trial (DCCT) was a 7-year longitudinal study that demonstrated
the importance of the intensive insulin therapy when compared to conventional treatment in the
development of chronic complications in patients with type 1 diabetes mellitus (T1DM). This study also
demonstrated another important issue: there is a reverse relationship between C-peptide levels
(endogenous indicator of insulin secretion) chronic complications - that is, the higher the C-peptide
levels, the lower the incidence of nephropathy, retinopathy and hypoglycemia. From such data, beta cell
preservation has become an additional target in the management ofT1DM [1].
Diabetology & Metabolic Syndrome 2009, 1:19 doi:10.1186/1758-5996-1-19
Carlos Eduardo Barra Couri* and Júlio César Voltarelli
Address: Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP,
Brazil Email: Carlos Eduardo Barra Couri* - ce.couri@yahoo.com.br; Júlio César Voltarelli - jcvoltar@fmrp.usp.br
* Corresponding author

15. 16
ABSTRACT
OBJECTIVE: To evaluate the effect of treatment of type 1 diabetes by transplantation of bone-derived stem
cells expressing human insulin gene.
METHODS: Murine bone marrow-derived stem cells expressing green fluorescent protein (GFP-mMSCs)
were isolated from 4/6-week-old GFP mice and transfected with a recombinant retrovirus-murine stem cell
virus (MSCV) encoding human insulin gene, thus constructing the GFP-mMSCs-MCV-insulin. 16 C57BL/6J
mice were injected with streptozotocin so as to establish models of type 1 diabetes and then randomly divided
into 4 equal groups: Group A, undergoing injection into the liver with GFP-mMSC-MCV-insulin 1 week after the
establishment of the model, Group B, undergoing intrahepatic transplantation of the GFP-mMSCs transfected
with blank vector, Group C, undergoing intrahepatic transplantation of untransfected GFP-mMSCs, and
Group D, undergoing intrahepatic transplantation of phosphate-buffered saline (PBS). Another 4 normal mice
were used as controls and underwent intrahepatic transplantation of PBS too. After the transplantation the
blood glucose, serum insulin, and body weight were detected everyday. 6 weeks later immunohistochemistry
was used to detect the expression of human insulin in the mice liver tissues.
RESULTS: The body weight of Group A increased by 6% within 6 weeks after treatment, and the average
blood glucose level 7 d and 42 d after transplantation were (10.4 +/- 2.8) mmol/L and (6.5 +/- 0.9) mmol/L
respectively, both significantly lower than those of Group D [(26.8 +/- 2.5) mmol/L and (25.4 +/- 4.1) mmol/L
respectively, both P < 0.05]. Immunohistochemistry showed secretion of human insulin in serum and liver.
CONCLUSION: The clinical manifestations of diabetes can be relieved effectively by intrahepatic
transplantation of mMSCs expressing human insulin gene. This study implies a novel approach of gene
therapy for type 1 diabetes.
Treatment of type 1 diabetes by transplantation of bone-derived mesenchymal
stem cells expressing human insulin gene: experiment with mice.
[Article in Chinese] Xu J, Zhu MY, LuYH, LuY, Wang ZW.
Source: Department of General Surgery,Affiliated Hospital of Nantong University, Nantong 226001, China.
Zhonghua Yi Xue Za Zhi. 2007 Sep 25;87(36):2557-60.
MesenchymalStemCellsforType1
DiabetesTreatment
28
Immunomodulation by Mesenchymal Stem Cells
A Potential Therapeutic Strategy for Type 1 Diabetes
Mesenchymal stem cells (MSCs) are pluripotent stromal cells that have the potential to give rise to cells of
diverse lineages. Interestingly, MSCs can be found in virtually all postnatal tissues. The main criteria currently
used to characterize and identify these cells are the capacity for self-renewal and differentiation into tissues of
mesodermal origin, combined with a lack in expression of certain hematopoietic molecules. Because of their
developmental plasticity, the notion of MSC-based therapeutic intervention has become an emerging
strategy for the replacement of injured tissues. MSCs have also been noted to possess the ability to impart
profound immunomodulatory effects in vivo. Indeed, some of the initial observations regarding MSC
protection from tissue injury once thought mediated by tissue regeneration may, in reality, result from
immunomodulation. Whereas the exact mechanisms underlying the immunomodulatory functions of MSC
remain largely unknown, these cells have been exploited in a variety of clinical trials aimed at reducing the
burden of immunemediated disease. This article focuses on recent advances that have broadened our
understanding of the immunomodulatory properties of MSC and provides insight as to their potential for
clinical use as a cell-based therapy for immune-mediated disorders and, in particular, type 1 diabetes.
Diabetes 57:1759–1767, 2008
RezaAbdi,1 Paolo Fiorina,1,2 Chaker N.Adra,1,3 MarkAtkinson,4 and Mohamed H. Sayegh1,3
From the 1Transplantation Research Center, Renal Division, Brigham and Women's Hospital and Children's Hospital Boston,
Harvard Medical School, Boston, Massachusetts; the 2Department of Medicine, San Raffaele Scientific Institute, Milan, Italy; the
3Stem Cell Therapy Program, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia; and the 4Department of
Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, Florida.
Corresponding author: RezaAbdi, rabdi@rics.bwh.harvard. DIABETES, VOL. 57, JULY2008
MesenchymalStemCellsforType1
DiabetesTreatment
BMC Med. 2012 Jan 10;10:3. doi: 10.1186/1741-7015-10-3.
29

16. 17
ABSTRACT
BACKGROUND: Inability to control autoimmunity is the primary barrier to developing a cure for type 1
diabetes (T1D). Evidence that human cord blood-derived multipotent stem cells (CB-SCs) can control
autoimmune responses by altering regulatory T cells (Tregs) and human islet â cell-specific T cell clones
offers promise for a new approach to overcome the autoimmunity underlying T1D.
METHODS: We developed a procedure for Stem Cell Educator therapy in which a patient's blood is circulated
through a closed-loop system that separates lymphocytes from the whole blood and briefly co-cultures them
with adherent CB-SCs before returning them to the patient's circulation. In an open-label, phase1/phase 2
study, patients (n=15) with T1D received one treatment with the Stem Cell Educator. Median age was 29 years
(range: 15 to 41), and median diabetic history was 8 years (range: 1 to 21).
RESULTS: Stem Cell Educator therapy was well tolerated in all participants with minimal pain from two
venipunctures and no adverse events. StemCell Educator therapy can markedly improve C-peptide levels,
reduce the median glycated hemoglobin A1C (HbA1C) values, and decrease the median daily dose of insulin
in patients with some residual â cell function (n=6) and patients with no residual pancreatic islet â cell function
(n=6).Treatment also produced an increase in basal and glucose-stimulated C-peptide levels through 40
weeks. However, participants in the Control Group (n=3) did not exhibit significant change at any follow-up.
Individuals who received Stem Cell Educator therapy exhibited increased expression of co-stimulating
molecules (specifically, CD28 and ICOS), increases in the number of CD4+CD25+Foxp3+Tregs, and
restoration of Th1/Th2/Th3 cytokine balance.
CONCLUSIONS: Stem Cell Educator therapy is safe, and in individuals with moderate or severe T1D, a single
treatment produces lasting improvement in metabolic control. Initial results indicate Stem Cell Educator
therapy reverses autoimmunity and promotes regeneration of islet âcells. Successful immune modulation by
CB-SCs and the resulting clinical improvement in patient status may have important implications for other
autoimmune and inflammation-related diseases without the safety and ethical concerns associated with
conventional stem cell-based approaches.
Reversal of type 1 diabetes via islet â cell regeneration following immune
modulation by cord blood-derived multipotent stem cells.
ZhaoY, Jiang Z, ZhaoT,Ye M, Hu C,Yin Z, Li H, ZhangY, DiaoY, LiY, ChenY, Sun X, Fisk MB, Skidgel R, Holterman M, Prabhakar B, MazzoneT.
Source: Section of Endocrinology, Diabetes & Metabolism, Department of Medicine, University of Illinois at Chicago, 1819 West Polk Street,
Chicago, IL60612, USA. yongzhao@uic.edu
MesenchymalStemCellsforType1DiabetesTreatment
30
ABSTRACT
The post-natal dental pulp tissue contains a population of multipotent mesenchymal progenitor cells known
as dental pulp stromal/stem cells (DPSCs), with high proliferative potential for self-renewal. In this
investigation, we explored the potential of DPSCs to differentiate into pancreatic cell lineage resembling islet-
like cell aggregates (ICAs). We isolated, propagated, and characterized DPSCs and demonstrated that these
could be differentiated into adipogenic, chondrogenic, and osteogenic lineage upon exposure to an
appropriate cocktail of differentiating agents. Using a three-step protocol reported previously by our group,
we succeeded in obtaining ICAs from DPSCs. The identity of ICAs was confirmed as islets by dithiozone-
positive staining, as well as by expression of C-peptide, Pdx-1, Pax4, Pax6, Ngn3, and Isl-1. There were
several-fold up-regulations of these transcription factors proportional to days of differentiation as compared
with undifferentiated DPSCs. Day 10 ICAs released insulin and C-peptide in a glucose-dependent manner,
exhibiting in vitro functionality. Our results demonstrated for the first time that DPSCs could be differentiated
into pancreatic cell lineage and offer an unconventional and non-controversial source of human tissue that
could be used for autologous stem cell therapy in diabetes.
Received August 23, 2010. Revision received December 1, 2010. Accepted December 1, 2010.
Differentiation of Dental Pulp Stem Cells into Islet-likeAggregates
V. Govindasamy1, V.S. Ronald2, A.N. Abdullah2, K.R. Ganesan Nathan1, Z.A.C. Ab. Aziz3, M. Abdullah3, S. Musa2, N.H.
Abu Kasim3#, and R. R. Bhonde1*
1
Stempeutics Research Malaysia Sdn Bhd, (773817-K), Lot G-E-2A, Enterprise 4, Technology Park Malaysia, Bukit Jalil, 57000
2
Kuala Lumpur, Malaysia Department of Children's Dentistry and Orthodontics, Faculty of Dentistry, University of Malaya, Kuala
3
Lumpur, Malaysia Department of Conservative Dentistry, Faculty of Dentistry, University of Malaya, 50603 Kuala Lumpur, Malaysia
* #
rrbhonde@stempeutics.com.my nhayaty@um.edu.my
Pleasemailthebulletpoint
31

17. 18
Abstract
Long-term studies strongly suggest that tight control of blood glucose can prevent the development and
retard the progression of chronic complications of type 1diabetes mellitus. In contrast to conventional insulin
treatment, replacement of a patient's islets of Langerhans either by pancreas organ transplantation of by
isolated islet transplantation is the only treatment to achieve a constant normoglycemic state and avoiding
hypoglycemic episodes, a typical adverse event of multiple daily insulin injections. However, the expense of
this benefit is still the need for immunosuppressive treatment of the recipient with all its potential risks. Islet cell
transplantation offers the advantage of being performed as a minimally invasive procedure, in which islets can
be perfused percutaneously into the liver via the portal vein. As of June 2003, 705 pancreatic islet transplants
worldwide have been reported to the International Islet Transplant Registry (ITR) at our Third Medical
Department, University of Giessen/Germany. Data analysis shows at 1year after adult islet transplantation a
patient survival rate of 97% a functioning islet graft in 54% of the cases, whereas insulin independence was
meanwhile achieved in 20% of the cases. However, using a novel protocol established by the Edmonton
Center/Canada, the insulin independence rates have improved significantly reaching meanwhile a 50-80%
level. Finally, the concept of islet cell or stem cell transplantation is most attractive since it offers many
perspectives: islet cell availability could become unlimited and islet or stem cells my be transplanted without
life-long immunosuppressive treatment of the recipient, just to mention 2 of them
Pancreatic islet and stem cell transplantation: new strategies in cell
therapy of diabetes mellitus.
Bretzel RG, Eckhard M, Brendel MD.Third Medical Department and Polyclinic, University Hospital Giessen, Giessen, Germany.
reinhard.bretzel@innere.med.uni-giessen.de
PMID:15238879 [PubMed - indexed for MEDLINE]
StemCellsforDiabetesCure
32
ABSTRACT
BACKGROUND: Cerebral palsy is currently one of the major diseases that cause severe paralysis of the nervous
system in children; approximately 9–30% of cerebral palsy patients are also visually impaired, for which no
effective treatment is available. Bone marrow mesenchymal stem cells (BMSCs) have very strong self-renewal,
proliferation, and pluripotent differentiation potentials. Therefore, autologous BMSC transplantation has become
a novel method for treating cerebral palsy.
METHODS: An 11-year-old boy had a clear history of dystocia and asphyxia after birth; at the age of 6 months, the
family members observed that his gaze roamed and noted that he displayed a lack of attention. A brain MRI
examination at the age of 7 years showed that the child had cerebral palsy with visual impairment (i.e., posterior
visual pathway injury). The patient was hospitalized for 20 days and was given four infusions of
intravenousautologous BMSCs. Before transplantation and 1, 6, and 12 months after transplantation, a visual
evoked potential test, an electrocardiogram, routine blood tests, and liver and kidney function tests were
performed.
RESULTS: The patient did not have any adverse reactions during hospitalization or postoperative follow-up. After
discharge, the patient could walk more smoothly than he could before transplantation; furthermore, his vision
significantly improved 6 months after transplantation, which was also supported by the electrophysiological
examinations.
CONCLUSIONS: The clinical application of BMSCs is effective for improving vision in a patient with cerebral palsy
combined with visual impairment.
Treatment of one case of cerebral palsy combined with posterior visual pathway
injury using autologous bone marrow Mesenchymal stem cells
Li et al. Journal ofTranslational Medicine 2012, 10:100 http://www.translational-medicine.com/content/10/1/100
Min Li1,AixueYu1, Fangfang Zhang1, GuangHui Dai1, Hongbin Cheng1, Xiaodong Wang1 andYihuaAn1,2*
StemCellTherapyinCerebralPalsyTreatment
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StemCellTherapyforParkinson's
DentalStemCellforNeuro
Regeneration
DentalStemCellforParkinson's
functionalrecovery
The progress of PD and its related disorders cannot be prevented with the medications available. In this study, we
recruited 8 PD and 4 PD plus patients between 5 to 15 years after diagnosis. All patients received BM-MSCs bilaterally
into the SVZ and were followed up for 12 months. PD patients after therapy reported a mean improvement of 17.92%
during “on” and 31.21% during “off” period on the UPDRS scoring system. None of the patients increased their
medication during the follow-up period. Subjectively, the patients reported clarity in speech, reduction in tremors,
rigidity, and freezing attacks. The results correlated with the duration of the disease. Those patients transplanted in
the early stages of the disease (less than 5 years) showed more improvement and no further disease progression
than the later stages (11–15 years). However, the PD plus patients did not show any change in their clinical status after
stem cell transplantation. This study demonstrates the safety of adult allogenic human BM-MSCs transplanted into
the SVZ of the brain and its efficacy in early-stage PD patients.
Bilateral Transplantation of Allogenic Adult Human Bone Marrow-Derived
Mesenchymal Stem Cells into the Subventricular Zone of Parkinson's Disease:APilot
Clinical Study
N. K. Venkataramana,1 Rakhi Pal,2 ShaileshA. V. Rao,1Arun L. Naik,1 Majahar Jan,2
Rahul Nair,2 C. C. Sanjeev,1 Ravindra B. Kamble,1 D. P.Murthy,1 and Krishna Chaitanya1
1Advanced Neuro Science Institute, BGS Global Hospital, Bangalore 560060, India. 2ANSA Research Foundation, Indiranagar, Bangalore
560038, India. Correspondence should be addressed to N. K. Venkataramana, drnkvr@gmail.com. Received 31 August 2011; Accepted 1
November 2011Academic Editor: Francesco Dazzi. Copyright © 2012 N. K. Venkataramana et al.
Volume 2012, Article ID 931902, 12 pages doi:10.1155/2012/931902
Human dental pulp-derived stem cells promote locomotor recovery after a complete
transection of the rat spinal cord by multiple neuro-regenerative mechanisms
Spinal cord injury (SCI) often leads to persistent functional deficits due to loss of neurons and glia and to limited
axonal regeneration after injury. Here we report that transplantation of human dental pulp stem cells intothe
completely transected adult rat spinal cord resulted in marked recovery of hind limb locomotor functions.
Transplantation of human bone marrow stromal cells or skin-derived fibroblasts led to substantially less recovery of
locomotor function. The human dental pulp stem cells exhibited three major neuroregenerative activities. First, they
inhibited the SCI-induced apoptosis of neurons, astrocytes, and oligodendrocytes, which improved the preservation
of neuronal filaments and myelin sheaths. Second, they promoted the regeneration of transected axons by directly
inhibiting multiple axon growth inhibitors, including chondroitin sulfate proteoglycan and myelin-associated
glycoprotein, via paracrine mechanisms. Last, they replaced lost cells by differentiating into mature oligodendrocytes
under the extreme conditions of SCI. Our data demonstrate that tooth-derived stem cells may provide therapeutic
benefits for treating SCI through both cell-autonomous and paracrine neuroregenerative activities.
Kiyoshi Sakai,1 Akihito Yamamoto,1 Kohki Matsubara,1 Shoko Nakamura,1 Mami Naruse,1 Mari Yamagata,1 Kazuma Sakamoto,2 Ryoji Tauchi,3
Norimitsu Wakao,3 Shiro Imagama,3 Hideharu Hibi,1 Kenji Kadomatsu,2 Naoki Ishiguro,3 and Minoru Ueda1 1Department of Oral and Maxillofacial
Surgery, 2Department of Biochemistry, and 3Department of Orthopedic Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan.
ABSTRACT
Human adult dental pulp stem cells (DPSCs), derived from third molar teeth, are multipotent and have the capacity to
differentiate into neurons under inductive conditions both in vitro and following transplantation into the avian embryo.
In this study, we demonstrate that the intracerebral transplantation of human DPSCs 24 hours following focal cerebral
ischemia in a rodent model resulted in significant improvement in forelimb sensorimotor function at 4 weeks post-
treatment. At this time, 2.3 ± 0.7% of engrafted cells had survived in the poststroke brain and demonstrated targeted
migration toward the stroke lesion. In the peri-infarct striatum, transplanted DPSCs differentiated into astrocytes in
preference to neurons. Our data suggest that the dominant mechanism of action underlying DPSC treatment that
resulted in enhanced functional recovery is unlikely to be due to neural replacement. Functional improvement is more
likely to be mediated through DPSC-dependent paracrine effects. This study provides preclinical evidence for the
future use of human DPSCs in cell therapy to improve outcome in stroke patients.
Human adult dental pulp stem cells enhance poststroke functional recovery through
non-neural replacement mechanisms.
Leong WK, HenshallTL,ArthurA, Kremer KL, Lewis MD, Helps SC, Field J, Hamilton-Bruce MA, Warming S, Manavis J, Vink R, Gronthos S, Koblar SA.
Source: Centre for Stem Cell Research, Robinson Institute, University ofAdelaide, SouthAustralia,Australia.
Stem Cells Transl Med. 2012 Mar;1(3):177-87. doi: 10.5966/sctm.2011-0039. Epub 2012 Feb 22.
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19. 09
MesenchymalStemCellsinOsteoarthitis
BACKGROUND: Osteoarthritis (OA) is a progressive disorder of the joints caused by gradual loss of articular
cartilage, which naturally possesses a limited regenerative capacity. In the present study, the potential of intra-
articular injection of mesenchymal stem cells (MSCs) has been evaluated in six osteoarthritic patients.
METHODS: Six female volunteers, average age of 54.56 years, with radiologic evidence of knee OA that
required joint replacement surgery were selected for this study. About 50 ml bone marrow was aspirated from
each patient and taken to the cell laboratory, where MSCs were isolated and characterized in terms of some
surface markers. About 20-24 × 10(6) passaged-2 cells were prepared and tested for microbial contamination
prior to intra-articular injection.
RESULTS: During a one-year follow-up period, we found no local or systemic adverse events. All patients were
partly satisfied with the results of the study. Pain, functional status of the knee, and walking distance tended to
be improved up to six months post-injection, after which pain appeared to be slightly increased and patients'
walking abilities slightly decreased. Comparison of magnetic resonance images (MRI) at baseline and six
months post-stem cell injection displayed an increase in cartilage thickness, extension of the repair tissue over
the subchondral bone and a considerable decrease in the size of edematous subchondral patches in three out
of six patients.
CONCLUSION: The results indicated satisfactory effects of intra-articular injection of MSCs in patients with
knee OA.
Intra-articular Injection of Autologous Mesenchymal Stem Cells in Six Patients
with Knee Osteoarthritis
Archives of Iranian Medicine, Volume 15, Number 7, July 2012
Mohsen Emadedin MD1, Naser Aghdami MD PhD1,2, Leila Taghiyar MSc2, Roghayeh Fazeli MD1, Reza Moghadasali MSc1,
Shahrbanoo Jahangir MSc2, Reza Farjad MD3
1Department of Regenerative Biomedicine and Cell Therapy, Cell Science Research Center, Royan Institute for Stem Cell Biology
andTechnology,ACECR,Tehran, Iran,2Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan
Institute for Stem Cell Biology andTechnology,ACECR,Tehran, Iran,
RolepfMesenchymalStemCellsin
Arthitistreatment
Mesenchymal stem cells in arthritic diseases
Mesenchymal stem cells (MSCs), the nonhematopoietic progenitor cells found in various adult tissues, are
characterized by their ease of isolation and their rapid growth in vitro while maintaining their differentiation
potential, allowing for extensive culture expansion to obtain large quantities suitable for therapeutic use. These
properties make MSCs an ideal candidate cell type as building blocks for tissue engineering efforts to
regenerate replacement tissues and repair damaged structures as encountered in various arthritic conditions.
Osteoarthritis (OA) is the most common arthritic condition and, like rheumatoid arthritis (RA), presents an
inflammatory environment with immunological involvement and this has been an enduring obstacle that can
potentially limit the use of cartilage tissue engineering. Recent advances in our understanding of the functions
of MSCs have shown that MSCs also possess potent immunosuppression and anti-inflammation effects. In
addition, through secretion of various soluble factors, MSCs can influence the local tissue environment and
exert protective effects with an end result of effectively stimulating regeneration in situ. This function of MSCs
can be exploited for their therapeutic application in degenerative joint diseases such as RA and OA. This
review surveys the advances made in the past decade which have led to our current understanding of stem cell
biology as relevant to diseases of the joint.
Faye H Chen and Rocky STuanArthritis Research &Therapy Vol 10 No 5
Cartilage Biology and Orthopaedics Branch,
National Institute ofArthritis, and Musculoskeletal and Skin Diseases, National Institutes of Health,
Department of Health and Human Services, Building 50, 50 South Dr., Bethesda, MD 20892, USA
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20. 21
Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-
versus-host disease: a phase II study.
ABSTRACT
BACKGROUND: Severe graft-versus-host disease (GVHD) is a life-threatening complication after allogeneic
transplantation with haemopoietic stemcells. Mesenchymal stem cells modulate immune responses in vitro and
in vivo. We aimed to assess whether mesenchymal stem cells could ameliorate GVHD after haemopoietic-stem-
cell transplantation.
METHODS: Patients with steroid-resistant, severe, acute GVHD were treated with mesenchymal stem cells,
derived with the European Group for Blood and Marrow Transplantation ex-vivo expansion procedure, in a
multicentre, phase II experimental study. We recorded response, transplantation-related deaths, and other
adverse events for up to 60 months' follow-up from infusion of the cells.
FINDINGS: Between October, 2001, and January, 2007, 55 patients were treated. The median dose of bone-
marrow derived mesenchymal stem cellswas 1.4x10(6) (min-max range 0.4-9x10(6)) cells per kg bodyweight. 27
patients received one dose, 22 received two doses, and six three to five doses of cells obtained from HLA-identical
sibling donors (n=5), haploidentical donors (n=18), and third-party HLA-mismatched donors (n=69). 30 patients
had a complete response and nine showed improvement. No patients had side-effects during or immediately
after infusions of mesenchymal stem cells. Response rate was not related to donor HLA-match. Three patients
had recurrent malignant disease and one developed de-novo acute myeloid leukaemia of recipient origin.
Complete responders had lower transplantation-related mortality 1 year after infusion than did patients with
partial or no response (11 [37%] of 30 vs 18 [72%] of 25; p=0.002) and higher overall survival 2 years after
haemopoietic-stem-cell transplantation (16 [53%] of 30 vs four [16%] of 25; p=0.018).
INTERPRETATION: Infusion of mesenchymal stem cells expanded in vitro, irrespective of the donor, might be an
effective therapy for patients with steroid-resistant, acute GVHD.
Le Blanc K, Frassoni F, Ball L, Locatelli F, Roelofs H, Lewis I, Lanino E, Sundberg B, Bernardo ME, Remberger M, Dini G, Egeler RM,
BacigalupoA, Fibbe W,Ringdén O; Developmental Committee of the European Group for Blood and MarrowTransplantation.
Source: Haematology Centre and Centre ofAllogeneic Stem Cell Transplantation, Division of Clinical Immunology, Karolinska University
Hospital, Huddinge, Stockholm, Sweden. katarina.leblanc@ki.se
Lancet. 2008 May 10;371(9624):1579-86. doi: 10.1016/S0140-6736(08)60690-X.
MesenchymalStemCellsfortreatmentofacutegraft-versus-
hostdisease
ABSTRACT
INTRODUCTION: Stem cells from human exfoliated deciduous teeth (SHED) have been identified as a
population of postnatal stem cells capable of differentiating into osteogenic and odontogenic cells, adipogenic
cells, and neural cells. Herein we have characterized mesenchymal stem cell properties of SHED in comparison
to human bone marrow mesenchymal stem cells (BMMSCs).
METHODS: We used in vitro stem cell analysis approaches, including flow cytometry, inductive differentiation,
telomerase activity, and Western blot analysis to assess multipotent differentiation of SHED and in vivo
implantation to assess tissue regeneration of SHED. In addition, we utilized systemic SHED transplantation to
treat systemic lupus erythematosus (SLE)-like MRL/lpr mice.
RESULTS: We found that SHED are capable of differentiating into osteogenic and adipogenic cells, expressing
mesenchymal surface molecules (STRO-1, CD146, SSEA4, CD73, CD105, and CD166), and activating multiple
signaling pathways, including TGFâ, ERK, Akt, Wnt, and PDGF. Recently, BMMSCs were shown to possess an
immunomodulatory function that leads to successful therapies for immune diseases. We examined the
immunomodulatory properties of SHED in comparison to BMMSCs and found that SHED had significant effects
on inhibiting T helper 17 (Th17) cells in vitro. Moreover, we found that SHED transplantation is capable of
effectively reversing SLE-associated disorders in MRL/lpr mice. At the cellular level, SHED transplantation
elevated the ratio of regulatory T cells (Tregs) via Th17 cells.
CONCLUSIONS: These data suggest that SHED are an accessible and feasible mesenchymal stem cell source
for treating immune disorders like SLE.
Immunomodulatory properties of stem cells from human exfoliated deciduous
teeth
TakayoshiYamaza1,2,#,Akiyama Kentaro1, #, Chider Chen1,Yi Liu1,Yufang Shi3, Stan Gronthos4, Songlin Wang5, Songtao Shi*
Yamaza et al. Stem Cell Research & Therapy 2010, 1:5 http://stemcellres.com/content/1/1/5
Immunomodulatorypropertiesof
DentalStemCells
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21. 22
MesenchymalStemCellsLowerIncidenceofAcuteRejectioninKidneyTransplantCases
Induction therapy with autologous mesenchymal stem cells in living-related
kidney transplants: a randomized controlled trial.
ABSTRACT
CONTEXT: Antibody-based induction therapy plus calcineurin inhibitors (CNIs) reduce acute rejection rates in
kidney recipients; however, opportunistic infections and toxic CNI effects remain challenging. Reportedly,
mesenchymal stem cells (MSCs) have successfully treated graft-vs-host disease.
OBJECTIVE: To assess autologous MSCs as replacement of antibody induction for patients with end-stage renal
disease who undergo ABO-compatible, cross-match-negative kidney transplants from a living-related donor.
DESIGN, SETTING, AND PATIENTS: One hundred fifty-nine patients were enrolled in this single-site,
prospective, open-label, randomized study from February 2008-May 2009, when recruitment was completed.
INTERVENTION: Patients were inoculated with marrow-derived autologous MSC (1-2 x 10(6)/kg) at kidney
reperfusion and two weeks later. Fifty-three patients received standard-dose and 52 patients received low-dose
CNIs (80% of standard); 51 patients in the control group received anti-IL-2 receptor antibody plus standard-dose
CNIs.
MAIN OUTCOME MEASURES: The primary measure was 1-year incidence of acute rejection and renal function
(estimated glomerular filtration rate [eGFR]); the secondary measure was patient and graft survival and incidence
of adverse events.
RESULTS: Patient and graft survival at 13 to 30 months was similar in all groups. After 6 months, 4 of 53 patients
(7.5%) in the autologous MSC plus standard-dose CNI group (95% CI, 0.4%-14.7%; P = .04) and 4 of 52 patients
(7.7%) in the low-dose group (95% CI, 0.5%-14.9%; P = .046) compared with 11 of 51 controls (21.6%; 95% CI,
10.5%-32.6%) had biopsy-confirmed acute rejection. None of the patients in either autologous MSC group had
glucorticoid-resistant rejection, whereas 4 patients (7.8%) in the control group did (95% CI, 0.6%-15.1%; overall P
= .02). Renal function recovered faster among both MSC groups showing increased eGFR levels during the first
month after surgery than the control group. Patients receiving standard-dose CNI had a mean difference of 6.2
mL/min per 1.73 m(2) (95% CI, 0.4-11.9; P=.04) and those in the low-dose CNI of 10.0 mL/min per 1.73 m(2) (95%
CI, 3.8-16.2; P=.002). Also, during the 1-year follow-up, combined analysis of MSC-treated groups revealed
significantly decreased risk of opportunistic infections than the control group (hazard ratio, 0.42; 95% CI, 0.20-
0.85, P=.02)
CONCLUSION: Among patients undergoing renal transplant, the use of autologous MSCs compared with anti-IL-
2 receptor antibody induction therapy resulted in lower incidence of acute rejection, decreased risk of
opportunistic infection, and better estimated renal function at 1 year.
Tan J, Wu W, Xu X, Liao L, Zheng F, Messinger S, Sun X, Chen J,Yang S, Cai J, Gao X, PileggiA, Ricordi C.
Source: Organ Transplant Institute, Fuzhou General Hospital, Xiamen University and Fujian Key Laboratory of Transplant Biology,
Fuzhou, China. tanjm156@yahoo.com.cn
JAMA. 2012 Mar 21;307(11):1169-77. doi: 10.1001/jama.2012.316.
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22. 23
Acute-on-chronic liver failure (ACLF) is a severe, life-threatening complication, and new and efficient therapeutic
strategies for liver failure are urgently needed. Mesenchymal stem cell (MSC) transfusions have been shown to
reverse fulminant hepatic failure in mice and to improve liver function in patients with end-stage liver diseases. We
assessed the safety and initial efficacy of umbilical cordderived MSC (UC-MSC) transfusions for ACLF patients
associated with hepatitis B virus (HBV) infection.A total of 43 ACLF patients were enrolled for this open-labeled
and controlled study; 24 patients were treated with UC-MSCs, and 19 patients were treated with saline as controls.
UC-MSC therapy was given three times at 4-week intervals. The liver function, adverse events, and survival rates
were evaluated during the 48-week or 72-week follow-up period. No significant side effects were observed during
the trial. The UC-MSC transfusions significantly increased the survival rates in ACLF patients; reduced the model
for end-stage liver disease scores; increased serum albumin, cholinesterase, and prothrombin activity; and
increased platelet counts. Serum total bilirubin and alanine aminotransferase levels were significantly decreased
after the UC-MSC transfusions. UC-MSC transfusions are safe in the clinic and may serve as a novel therapeutic
approach for HBV-associated ACLF patients.
Human Mesenchymal Stem Cell Transfusion Is Safe and Improves Liver Function
inAcute-on-Chronic Liver Failure Patients
MING SHI,a,b* ZHENG ZHANG,a* RUONAN XU,a* HU LIN,a JUNLIANG FU,a ZHENGSHENG ZOU,a
AIMIN ZHANG,a JIANFEI SHI,a LIMING CHEN,a SALV,aWEIPING HE,a HUAGENG,a LEI JIN,a
ZHENWEN LIU,b FU-SHENGWANGa
Key Words. Clinical trials Stem cell transplantation Umbilical cord Liver
aResearch Center for BiologicalTherapy, Institute ofTranslational Hepatology,
and bResearch Center for LiverTransplantation, Beijing 302 Hospital, Beijing, China
MesenchymalStemCellsforLiverFailure
Treatment
High-purity hepatic differentiated from dental pulp stem cells in serum-free
medium
INTRODUCTION: We have previously differentiated hepatocyte like cells from deciduoud tooth pulp stem and
extracted third molar pulp stem cells with a protocol that used fetal bovine serum, but it showed high
contaminations of nondifferentiated cells. Both the lower purity of hepatically differentiated cells and usage of
serum are obstacles for applications of cell therapy or regenerative medicine. Objective of this study was to
investigate the capacity for and purity of hepatocyte-like differentiation of CD 117-positive dental pulp stem cells
without serum.
METHODS: Mesenchymal cells from human deciduous and extracted third molar pulp were isolated and
expanded in vitro. We separated D117-position cells by using a magnetic-activated cell sorter. The cells were
characterized immunofluorescently by using known stem cell markers. For hepatic differentiation, the media were
supplemented with hepatic growth factor, insulin transferring-selenium-x, dexamethasone, and oncostatin M.
Expression of hepatic markers alpha fetoprotein, albumin, hepatic nuclear factor-4 alpha, insulin-like growth
factor-1, and carbamoyl phosphate synthetase was examined immunoflurescently after differentiation. The
amount of differentiated cells was assessed by using flow cytometry. Glycogen storage and area concentration in
the medium were defined.
RESULTS: Both cell cultures demonstrated a number of cells positive for all tested hepatic markers after
differentiation,ie, albumin-positive cells were almost 90% of differentiated deciduous pulp cells. The
concentration of urea in the media increased significantly after differentiation. Significant amount of cytoplasmic
glycogen storage was found in the cells.
CONCLUSIONS: Without serum both cell types differentiated into high-purity hepatocyte-like cells. These cells
offer a source for hepatocypte lineage differentiation for transplantation in the future.
J Endod 2012 Apr;38(4):475-80. Epub 2012 Jan28
DentalStemCellsConverttoHepaticCells
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42

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