2. The purpose of this session is to provide possibilities in stem
cell therapy and regenerative medicine including their
limitations.
The session should not be interpretation out of context
The contents used are solely for understanding of the subject
on the basis of evidence based facts.
DISCLAIMER
3. “If the potential of stem cell research is
realized, it would mean an end to the
suffering of millions of people. If stem cell
research succeeds, there isn’t a person in
the country who won’t benefit, or know
someone who will”
- Michael J. Fox
4. God grant me the serenity to accept
things I cannot change
Courage to change the things I can
And wisdom to know the difference
Reinhold Niebuhr
A WISE SAYING
5. Autoimmune diseases - modifying the immune system
Diabetes - restoring pancreatic function
Oncology and cancer treatment
Cardiovascular diseases - healing the heart and vascular system
Neurological and cerebrovascular disorders - regenerating CNS tissue
Ophthalmic disorders - potential for overcoming sight loss
Bioartificial livers and hepatic regeneration with stem and progenitor cells
Dermatology - developments in cellular skin repair
Adipose-derived stem cells for autologous treatments
Regenerative Dentistry - potential for biotechnological tooth replacement.
Thrust Areas for Regenerative Therapy
9. Regenerative Applications – Blood Disorders
Aplastic Anemia
Fanconi Anemia (The first cord blood transplant in 1988 was for FA, an inherited disorder)
Sickle Cell Disease
Beta Thalassemia Major & Minor
SCID with Adenosine Deaminase Deficiency (ADA-SCID)
SCID which is X-linked (Severe Combined Immuno Deficiency)
SCID with absence of T & B Cells
SCID with absence of T Cells, Normal B Cells
Ataxia-Telangiectasia
Lymphoproliferative Disorders
Myeloproliferative Disorders
Acute Myelofibrosis
Agnogenic Myeloid Metaplasia (Myelofibrosis)
Polycythemia Vera
Essential Thrombocythemia
Chronic Granulomatous Disease
Neutrophil Actin Deficiency
Reticular Dysgenesis
Bone Marrow Cancers
Multiple Myeloma
Plasma Cell Leukemia
10. Therapy
- Dendritic Cells (DC) therapy – Only Autologous
- Allogeneic Natural Killer (NK) Cell Therapy
- Allogeneic Cytotoxic T Lymphocyte Therapy
Blood cancers or Solid Cancers
Intervention Model ( Therapy – NK cell Therapy); Trials ( DC therapy)
Primary Purpose: Treatment; Status : Approved
Advanced Cancer Therapies
CAR T Cell Therapy
Cancer Immunotherapy
11. CASE OF SPINAL CORD INJURY
Accident-2011
Damage-lower back and hips
On Bed/Wheelchair-2011-2017
Treatment –Stem cell transplantation
(UCB) 2017
26. Schmelzetheisen et al.first showed feasibility of using a
tissue engineered bone graft formed by periosteum –
derived stem cells for augmentation in the posterior
maxilla prior to impalnt insertion
TISSUE ENGINEERED BONE
GRAFT
27. Direct use of patient derived fresh cellular graft prepared
at the chair side .These procedures are relatively
convenient for clinicians because they do not involve lab
intervention. In 2006 ,Smiler and Soltan reported a
technique for chair side cellular graft preparation using
fresh aspirated bone marrow from the ileum that was
mixed with biocompatible matrix and scaffolds
Active Motif's Stem Cell CDy1 Dye
AFTER 3 MONTHS
32. HIGH PLASTICITY
CRYO-PRESERVED FOR A LONGER PERIOD
GOOD INTERACTION WITH
SCAFFOLD
GROWTH FACTORS
DENTAL STEM CELL ADVANTAGE
33. MSCs can simply be isolated by adherence to cell
culture plastic. The number of mononuclear cells in the
sample is determined and seeded at a density of 5000
cells/cm2.
ISOLATION
36. “In the fields of observation chance favors only the prepared
mind”
Pasteur
• Curosity driven- Alexande Fleming/ Joseph Mering and
Oscar minkowski
• Need driven
• Profit driven
• Opportunity driven
STEM CELLS RESEARCH
4/14/2017
7/26/2017
37.
38. TISSUE ENGINEERING FOR TMJ
Scaffold
Degradation=Matrix Synthesis
Cells
Source
Seeding Density
GF’s
PDGF-AB
bFGF
IGF-1
40. SCAFFOLD FREE
Hu JC and Athanasiou KA (2006).A self-assembling
process in articular cartilage tissue engineering.
Tissue Eng. 2006 Apr;12(4):969-79.
Stem cells have tremendous promise to help us understand and treat a range of diseases, injuries and other health-related conditions. Their potential is evident in the use of HSC to treat diseases of the blood, a therapy that has saved the lives of thousands of children with leukemia; and can be seen in the use of stem cells for tissue grafts to treat diseases or injury to the bone, skin and surface of the eye. Important clinical trials involving stem cells are underway for many other conditions and researchers continue to explore new avenues using stem cells in medicine. Stem cells could lead to breakthroughs in treatments and cures for any terminal or catastrophic disease you can think of .this is one of the reasons that support for this work has galvanized a coalition of advocates from just about every patient community.
The patients lower limb were paralysed after an accident in 2011.damaged her lower back and hips .since then spent her life in bed or a wheel chair.The stem cell transplantation (Umbilical cord blood) was done in 2017 and in just 2 months,she started walking with the help of a walker under supervision
The ultimate goal for tissue engineering and regenerative medicine is to develop therapies to restore lost, damaged, or aging tissues using engineered or regenerated products derived from either donor or autologous cells. Cell-based therapies are the most common approaches in regenerative medicine. Challenges in applying this approach clinically are to acquire the appropriate source of cells, to identify methodologies to induce cell proliferation and differentiation, to maintain cell survival, and to remove unwanted cells.
Stem cell plasticity is the ability of cells of a given type to be biochemically prompted grow into a different type of cell, whether the difference is modest (such as one kind white blood cellbeing used to grow another kind) or pronounced (such as a cell taken from the umbilical cord being used to create heart tissue
For the first few isolates it is advised to screen isolated cells for MSC markers (for instance testing positive for CD29, CD44, CD105 and CD166) to check whether you got the right cells. You can do a density centrifugation step before this using for instance Ficoll, but in my experience this is not absolutely necessary.
Expansion of hematopoietic stem cells (HSCs) has remained an important goal to develop advanced cell therapies for blood disorders. During the last 2 decades, ever since the first hematopoietic growth factors were identified and isolated, there have been serious attempts to expand HSCs in vitro using purified growth factors that are known to regulate primitive hematopoietic cells. However, these attempts have met with limited success. For example, the hematopoietic growth factors fetal liver tyrosine kinase (Flt3) ligand, stem cell factor, and interleukins 6 and 11 promoted self-renewal of murine hematopoietic stem cells, although only a limited expansion of stem cells compared with fresh input cells could be obtained.1 More recently, regulatory pathways other than those activated by the traditional hematopoietic growth factors have been investigated.
Alexander Fleming in the summer of 1928 was not looking for an antibacterial agent at the time a spore floated into his petridish.But he was extremely well read and trained in microbiology and could easily recognize the meaning of the clear area in the bacterial culture produced by the accidental implantation of the mould
In 1889,while studying the function of the pancreas in digestion, Joseph Mering and Oscar minkowski removed the pancreas from a dog. One day later, a laboratory assistant called their attention to a swarn of flies around the urine from the dog .Curious about why the flies were attracted to the urine, they analyzed it and found it was loaded with sugar, a common sign of diabetes. Later in 1921 Fredrick Bantin etal proved effectiveness of pancreatic secretion in diabetes
Stem cells are unique in many ways. While they present several potential clinical benefits
as reported through controlled clinical trials, there are equally unforeseen hazards for their
Use.However, the biological properties of these cells and the effect of their processing
and ex vivo handling raise specific concerns. Of these major are specific to their collection,
processing, storage and use for clinical applications. It must be understood that the donor
has the exclusive right to get apprised of all details related to his/her health and safety.
Tissue engineering is emerging as a promising option to repair or, potentially replace the diseased tissues of the mechanically demanding, and biologically complex TMJ. Traditionally, the principal elements of tissue engineering are cells, stimuli, and scaffolds .
Scaffold-based approaches
Scaffolds serve as a supportive structure to render shape and volume to the engineered tissues, allowing for well-defined geometries and ease of handling [70]. The scaffold diffusional characteristics can be modulated by changing pore size and porosity. The mechanical properties and surface characteristics can also be tailored to the tissue being engineered.
Scaffold degradation rates depend on scaffold chemistry and can be altered by manipulating
Central to the efforts of tissue engineering are identifying a suitable source of cells and seeding density. TMJ disc cells, as well as articular chondrocytes derived from the condyle, fossa-eminence and shoulder, in addition to dermal fibroblasts have all been examined for engineering the TMJ disc [81]. For example, dermal fibroblasts showed chondrogenic potential when treated with IGF-1. These cells are particularly promising since they are clinically relevant for autologous therapy without significant donor site morbidity. Cell seeding density is another important factor that influences the composition of the resultant construct [82]. Increasing cell seeding density does not always improve functional or biomechanical properties, so seeding density must be carefully controlled. For example, TMJ disc cells seeded from 15-120M cells/ml of scaffold volume onto PGA scaffolds show variable results. Increasing the cell number up to 120M cells/ml of scaffold volume increased GAG and collagen content without significant improvement in the compressive properties of the engineered tissue. For each new cell source, identified for engineering the TMJ disc, the lowest seeding density that yields desirable functional properties must be determined.
Bioactive signals have been used to promote collagen and GAG synthesis in engineered TMJ discs, with the expectation that this approach would lead to improved mechanical properties of the engineered tissues. Effects elicited by bioactive signals depend greatly on conditions of the experiment. For example, in a study that compared platelet derived growth factor-AB (PDGF), bFGF, and IGF-1 at various concentrations in monolayer, bFGF was shown to result in the greatest improvements in GAG and collagen production (∼2 and 4.5 fold increases, respectively, compared to control) [86]. However, when examining IGF-1, bFGF, and TGF-β1 on cell-seeded PGA scaffolds in spinner flasks, the results suggested that IGF-1 elicited the greatest collagen production [73]. Both studies used porcine TMJ disc cells, but the conditions varied, the former was a two-dimensional, static culture, while the latter was a three-dimensional culture subjected to fluid-induced shear. Due to the limited number of studies on the efficacy of growth factors for scaffold-based engineering of TMJ discs, additional work is needed to optimize relevant growth factors, doses and regimens. However, a comprehensive picture will not emerge until this characterization is performed for each scaffold and culture condition.
As noted above, scaffold characteristics must be tailored to the cell types used to engineer the tissue. For example, porcine TMJ disc cells seeded on PGA scaffolds result in constructs that contract severely because the PGA degrades much faster than the matrix is produced. Although the addition of insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (bFGF), and transforming growth factor-β1 (TGF-β1) improved matrix synthesis [73, 74], the scaffold still degradesfast. As a result, a different scaffold material, PLLA, was examined for its much slower degradation rate [75]. Overall, PLLA scaffolds seeded with porcine TMJ cells and treated with TGF-β1 demonstrated higher collagen and GAG contents and improved mechanical properties (1.4 MPa Young's modulus) when compared to constructs seeded on PGA [75].
The advent of new manufacturing techniques may allow for the production of scaffolds that more closely mimic the unique structures of TMJ components, including tissue anisotropy. Toward this end, additive manufacturing was used to 3D print PCL scaffolds with an anisotropic internal structure [79••]. Seeded with mesenchymal stem cells (MSCs), anisotropic properties were observed. However, given the short time points examined in this study, it is unclear if the anisotropic properties would be retained long-term, since the observed anisotropy was likely due to the scaffold as opposed to the matrix produced. The goal of using scaffolds, to engineer anisotropic TMJ tissues composed of only cell-generated matrices, is yet to be realized.
-----agarose in vitro without using a scaffold
There is still a lot to learn about stem cells, however, their current applications as treatments are sometimes exaggerated by the media and other parties who do not fully understand the science and current limitations, and also by “clinics” looking to capitalize on the hype by selling treatments to chronically ill or seriously injured patients.
