This study compared genes related to cardiovascular development in stem cells from dental pulp and adipose tissue. Both stem cell types showed stem cell morphology and could differentiate into osteoblasts, adipocytes, and chondroblasts. A human stem cell transcription factor array showed dental pulp stem cells expressed more pluripotency markers while adipose stem cells expressed more obesity-related genes. Both stem cell types were prone to angiogenesis when differentiated into cardiomyocytes, but adipose stem cells expressed more cardiac biomarkers. Neither stem cell type showed spontaneous beating, indicating incomplete cardiomyocyte differentiation. The study aimed to identify suitable stem cell sources for cardiac regeneration.
Okyanos Heart Institute Stem Cell Therapy Educational SeminarErika Rosenthal
Okyanos Heart Institute treats patients with coronary artery disease with their own stem cells. This presentation is part of a free educational seminar. More information can be found at http://www.okyanos.com
The collection and processing of hematopoieticakshaya tomar
BASICS OF HSC COLLECTION AND PROCESSING INCLUDING ALL THE THREE SOURCES, A BRIEF ABOUT STEM CELL MOBILIZATION, STEM CELL SELECTION CRYOPRESERVATION AND DMSO
Okyanos Heart Institute Stem Cell Therapy Educational SeminarErika Rosenthal
Okyanos Heart Institute treats patients with coronary artery disease with their own stem cells. This presentation is part of a free educational seminar. More information can be found at http://www.okyanos.com
The collection and processing of hematopoieticakshaya tomar
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Effect of stemregenin1 and sb431542 small molecules on ex vivo expansion of u...Liberty University (LU)
Cord blood Hematopoietic stem cells (HSCs) with several advantages including low chance of viral contamination and low rate of Graft versus host disease (GVHD) are appropriate candidate for vast medical applications such as transplantation. The main obstacle of cord blood HSCs is the low number cells. To improve ex-vivo expansion of umbilical cord HSCs we introduced a new culture system. Isolated HSCs were seeded in Three-dimensional(3D) on Polyethersulfone(PES) scaffolds and Twodimensional(2D) culture conditions and treated with SB431542 and Stemregenin1(SR1) small molecules. On the fifth and tenth days the expanded cells in different groups were investigated for expression of specific markers by flow cytometry, expression of some stemness genes by qRT-PCR and colony formation by methocult medium. SR1 molecule significantly increased expansion of CD34+ cells while SB431542 induced more CD34+/38+ cells. Also SB431542 treated cells showed higher colony formation capacity. SR1 increased the expression of c-Myc, HOXB4 and SALL4 while SB431542 seemed to inhibit HOXB4 expression and increase SALL4.Together all, this study introduced a new ex vivo culture setting for further medical application of HSCs. Our data showed simultaneous use of these two small molecules can provide appropriate outcome for HSCs transplantation includes both of engraftment and repopulation.
ABSTRACT- Coronary artery disease (CAD) is suspected as a leading cause of mortality in developed countries. Due
to cholesterol and fat deposit plaque is forming into the inner walls of the arteries of the heart, which leads to narrowing
of blood vessels of heart and reduce the blood flow rate into heart. Proprotein convertase subtilisin-like kexin type 9
(PCSK9) is one of the candidate gene that regulate lipoprotein retention pathway of CAD development. It is a newly
discovered serine protease that plays a key role in LDL-C homeostasis by mediating LDL receptor (LDLR). The LDL
receptor is breakdown through a post transcriptional mechanism and induces the production of very low-density
lipoprotein in the fasting state. The aim of this study was to investigate the frequency of single nucleotide
polymorphism (SNP) of PCSK9 gene of 155 CAD patients and 102 ages matched healthy controls. Serum lipids
including total cholesterol (TC), triglycerides (TG), HDL, LDL, and VLDL were analyzed. PCR-RFLP analysis was
carried out to genotype regions carrying Eam 1104I restriction site in the PCSK9. Gene considering significant
difference in serum TC, TG, HDL-C, LDL-C and VLDL-C levels (P<0.001, <0.0001) of patients and control samples.
In CAD patients, G allele frequency is less than A allele frequency. G allele is responsible for decreasing the
LDL: HDL ratio which shows evidence in having its protecting effect on the occurrence of CAD in West Bengal Population.
Key-words- CAD, PCSK9, SNP, Eam1104I, Polymorphism, West Bengal population
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
Acorn Recovery: Restore IT infra within minutesIP ServerOne
Introducing Acorn Recovery as a Service, a simple, fast, and secure managed disaster recovery (DRaaS) by IP ServerOne. A DR solution that helps restore your IT infra within minutes.
This presentation by Morris Kleiner (University of Minnesota), was made during the discussion “Competition and Regulation in Professions and Occupations” held at the Working Party No. 2 on Competition and Regulation on 10 June 2024. More papers and presentations on the topic can be found out at oe.cd/crps.
This presentation was uploaded with the author’s consent.
Have you ever wondered how search works while visiting an e-commerce site, internal website, or searching through other types of online resources? Look no further than this informative session on the ways that taxonomies help end-users navigate the internet! Hear from taxonomists and other information professionals who have first-hand experience creating and working with taxonomies that aid in navigation, search, and discovery across a range of disciplines.
0x01 - Newton's Third Law: Static vs. Dynamic AbusersOWASP Beja
f you offer a service on the web, odds are that someone will abuse it. Be it an API, a SaaS, a PaaS, or even a static website, someone somewhere will try to figure out a way to use it to their own needs. In this talk we'll compare measures that are effective against static attackers and how to battle a dynamic attacker who adapts to your counter-measures.
About the Speaker
===============
Diogo Sousa, Engineering Manager @ Canonical
An opinionated individual with an interest in cryptography and its intersection with secure software development.
Sharpen existing tools or get a new toolbox? Contemporary cluster initiatives...Orkestra
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James Wilson, Orkestra and Deusto Business School
Emily Wise, Lund University
Madeline Smith, The Glasgow School of Art
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Grds 26 dec 2014 edited
1. Comparative analysis of cardiovascular development
related genes in stem cells isolated from
deciduous pulp and adipose tissue
Loo Zhang Xin
(Phd candidate)
ICAET, 26-27 December 2014
3. Cell therapy as an alternative?
• Left ventricular of the human heart contains ~5.8 x 109
cardiomyocytes.
• Upon the onset of AMI, 25% of the cardiomyocytes will be lost.
• Regeneration of the infarcted heart potentially requires:
– a billion cardiomyocytes,
– along with other supporting cells such as endothelial and smooth vascular cells
for functional integration.
• Possible few mechanisms for which these cells may influence the cardiovascular
regeneration:
– Differentiation of stem cells into cardiomyoctes,
– Fusion of stem cells with endogenous cells,
– Stem cells secretome signaling
4. List of suitable candidates
Embryonic stem cells
(ESC)
Induced pluripotetent
stem cells (IPS)
Adult stem cells
Pros • Totipotency
• Laflamme et al.
showsed that ESC were
able to generate
cardiomyocytes
and had limited death
rate in animal model.
• Can generate a
patient’s specific stem
cells.
• Ieda et al. used a
combination of cardiac
transcription factors to
direct reprogram
cardiac fibroblast into
cardiomyocytes.
• Cardiac stem cells
(CSCs) offers better
prospects.
• Bolli et al. reported
successful clinical trial
using CSCs in human
subjects with ischemic
cardiomyopathy had
been reported.
Cons • Ethical issues
destruction of embryo
• Complicated isolation
methods
• Tendency to form
tumours
• Risk of insertional
mutagenesis is of
particular concern.
•Invasive procedures in
isolating and culturing
the cells
• Escalating production
cost due to autologous
settings.
6. Methodology
• Characterized MSCs sources: human dental pulp (SHED) and human
adipose stem cells (ASC)
Morphology, Tri-lineage differentiation and immunophenotyping
Human Stem Cell Transcription Factors Array for initial screening
Ingenuity Pathway Analysis
• Cardiac differentiation using defined media
SHED and ASC were seeded onto 6-well plates at a density of 100,000 cells/cm2
.
Cardiac media: KO-DMEM containing 0.5% penicillin/streptomycin and
1% 1X Glutamax were supplemented with 100 ng/ml human recombinant Activin A
for 24 hours followed by 10 ng/ml human recombinant BMP2 for 4 days.
The medium was then exchanged for KO-DMEM without supplementary cytokines,
and cultures were refed every 2–3 days in a humidified incubator at 37°C and 5 %
CO2 for up to 14 days.
Cell morphology was captured using an inverted microscope, RNA was extracted at
day 7 and day 14, immunocytochemisty at day 14.
9. MSCs derived from SHED
and ASC:
Fibroblastic morphology
Expressed MSCs markers
Capable of differentiating
into osteoblasts, adipocytes
and chondroblasts
10. WHY Human stem cell
transcription factor array ?
REASON: Plenty of
experimental works
emphasized on
understanding stem cell
characteristics such as
proliferation pathways,
quality of the cells under in
vitro culture conditions &
lack of information
pertaining to stem cells at the
basal level.
Observation:
Higher expression level for
the HOX group of genes in
the ASC [Development of
obesity and body fat
distribution]
SHED expressed many
markers related to
pluripotency [More primitive
11.
12. • Both types of cells are prone to angiogenesis as there are more
interconnected molecules involved in formation of blood vessels and
vasculogenesis.
• This provides an explanation of how stem cells are able to improve or
preserve cardiac function in an animal model through angiogenesis and not
due to cardiomyocyte differentiation.
• In hypoxic conditions, expression of HIF-α increases rapidly which in turn
activates genes that encode for pro-angiogenic growth factors, such as
VEGF, BGFG, Ang-1 and 2, PFG and PDGF-B.
• These released cytokines encourage angiogenesis and the angiogenic effects
of human multipotent stromal cell which explains the underlying
improvement of cardiac function through any individual or a combination of
mechanisms such as apoptosis inhibition, increase in survival, and
angiogenesis stimulation by activation of the PI3K-Akt pathway.
13. ASC: Assumed more of a polygonal
shape.
SHED: Elongated and irregular
There has been no reports yet of a
successful protocol using dental stem
cells to generate beating cardiomyocytes.
Prior examples utilized dental pulp
stem cells with a combination of VEGF,
BFGF and IGF-1.
In this case, spontaneous cellular
beating was also not detected and the
morphology similar to what we observed
in our experiments.
Only GATA4 and NKX 2.5 were
detected in differentiated ASC
14. ASC expressed significantly more
cardiomyocyte biomarkers compared
to SHED at both day 7 and day 14
[Could be both adipose and heart
development are mesoderm origin.
This indicates that not all sources of
MSCs are suitable for cardiac
differentiation & selection of the MSC
source for treating a disease highly
depended on its origin.
Spontaneous cellular beating, the
hallmark indicator of successful
cardiomyocyte differentiation was not
detected in our experiment
[Could be cTnI was suppressed in this
condition whereby it can only be
activated when the cells were cultured
with neonatal rat cardiomyocytes, or
when these cells were delivered in
vivo to murine models of myocardial
infarction]
15. Acknowledgement
This work is part of a research collaboration between Hygieia
Innovation and the Faculty of Dentistry, University of Malaya and is
supported by the University of Malaya, High Impact Research Grant,
Ministry of Higher Education, Malaysia.
(Grant No UM.C/HIR/MOHE/DENT/02)