PIONEERING R & D; NURTURING COLLABORATION

Recognised Research Center Of Mysore University

Pre-clinical Studies


Comparitive evaluation of mesenchymal stem cells derived from bone marrow and Wharton’s jelly of umbilical cord for the treatment of type 1 diabetes mellitus: A pre clinical study
  • Type 1 diabetes mellitus is an autoimmune disorder where in the body recognises the pancreatic islet cells as foreign, resulting in the destruction of the insulin producing cells. This debilitating condition makes the patient in question resort to continuous supplementation of insulin to take the life further. The current therapeutic modules are only supporting in nature without catering to cure the cause of the disease. This made us create an animal model of type 1 DM and to evaluate the efficacy of mesenchymal stem cells derived from bone marrow and Wharton’s jelly of umbilical cord for the treatment of type 1 diabetes mellitus. Experimental diabetes was induced in rats by intra peritoneal administration of Streptozotocin (STZ). After 7 days blood glucose levels were measured & rats with glucose levels above 200 mg/dl were selected as diabetic rats & included in experiment. STZ induced diabetic rats were divided into different groups receiving mesenchymal stem cells derived from bone marrow and Wharton’s jelly of umbilical cord, Standard of care – Insulin and Glybenclamide and the control animals. Clinical parameters of recovery like reduction in blood glucose level, C Peptide level, body weight and water intake post treatment were evaluated. Regular histology as well as immunohistochemistry of the pancreatic samples obtained from study groups was also performed. The current study shown the supremacy of mesenchymal stem cells derived from Wharton’s jelly of umbilical cord as a promising tool which ensures the regeneration of destroyed islets.

  • The histological evidence in all the groups – pre and post transplantation is concordant with the results obtained in the serum biochemistry study as well as the clinical outcome. In diabetic animals which have received WJ MSC’s shows extensive regeneration in terms of normalof pancreatic lobes, acini and the presence of near normal regenerated islet cells. Whereas, histologically no regenerative or reparative changes were observed in diabetic animals which received BM MSC’s, Insulin and Glybenclamide. And their histogram was more similar to positive diabetes control pancreatic tissue sample.

Bone marrow mesenchymal stem cells as a promising candidate for treatment of spinal cord injury : A pre clinical study
  • Spinal cord injury (SCI) is a medically untreatable condition for which stem cells have created hope. Pre-clinical and clinical studies have established that these cells are safe for The dose dependency, survivability, route of administration, cell migration to injury site and effect on sensory and motor behavior in an SCI-induced paraplegic model were studied. A spinal cord contusion injury model was established in rats. Bone marrow (BM) mesenchymal stromal cells (MSC) were tagged to facilitate tracing in vivo. Two different doses (2 and 5 cells/kg body weight) and two different routes of infusion (site of injury and lumbar ) were tested during and after the spinal shock period. The animals were tested post-transplantation for locomotor capacity, motor control, sensory reflex, posture and body Stem cell migration was observed 1 month post-transplantation in spinal cord sections. The overall results demonstrated that transplantation of BM MSC significantly improved the locomotor and sensory behavior score in the experimental group compared with the group, and these results were dose dependent. All the infused stem cells could be visualized at the site of injury and none was visualized at the injected site. This indicated that the cells had survived in vivo, were probably chemoattracted and had migrated to the lesion site. To conclude, MSC transplanted with a lumbar puncture method migrate to the site of injury and are the most suitable for SCI healing. These cells demonstrate a dose-dependent effect and promote functional recovery when injected during or after the spinal shock period.

  • Comparative evaluation of mesenchymal stem cells derived from bone marrow and Wharton’s jelly of umbilical cord for the treatment of rheumatoid arthritis: A pre clinical study
    • Rheumatoid arthritis is a chronic auto immune disorder which primarily affects the joints. The immune system of the body identifies the cartilaginous matrix of the articular capsule as thereby causing a higher influx of inflammatory cells. This eventually results in an immune mediated destruction of the affected joint. Though a number of modalities are in place for the treatment of rheumatoid arthritis, one that addresses the basic etiology is still lacking. The well studied and proven immunomodulatory properties of mesenchymal stem cells lead us to perform a pre clinical trial to assess the comparative efficacy of mesenchymal stem cells from bone marrow and Wharton’s jelly of umbilical cord for the treatment of rheumatoid arthritis. We could successfully create a pre clinical model mimicking auto immune arthritis. Collagen induced auto immune arthritic Wistar rats were used in the study. Physical parameters like swelling and pain index, biochemical parameter viz. IL1 beta and histopathological changes of the joints were analyzed pre and post therapy. All the parameters suggested the comparative supremacy of mesenchymal stem cells derived from Wharton’s jelly of umbilical cord for the treatment of rheumatoid arthritis.

    Trans - Differentiation of Human Perinephric Fat Mesenchymal Stem Cells (hPNF-MSCs) into Endodermal and Mesodermal lineages - a future for cell based therapeutics
    • Stem cell populations are established within "niches"- specific anatomic locations that regulate how they participate in tissue regeneration, maintenance and repair. Each organ in the adult human body has its own niche of stem cells known as mesenchymal stem cells. Growing inter-est in mesenchymal stem cell based cell therapy made researchers to isolate these cells from a variety of tissue sources. However the supply of adult tissues is limited due to inadequate number of living donor participation. Adipose tissue is extensively explored for isolation of mesenchymal stem cells since ADSCs are acquired from waste product obtained during . Regenerative medicine challenges researchers to find non- controversial, safe and abundant stem cell sources.

    • In this context, we attempted to isolate and characterize mesenchymal stem cell population from deceased donor derived human perinephric fat in terms of morphology, growth kinetics and immunophenotype. hPNF-MSCs were isolated from deceased donors with informed consent from the legal representative and propagated in- vitro till passage 4. Cells showed fibroblast like morphology at P0 with persistent expression of MSC surface markers- CD 44, CD 73, CD 90, CD 105 and CD 166 propagation fulfilling the ISCT criteria for mesenchymal stem cells. Results obtained from the study were compared with that of previous study carried out using PNF from live donor. Live donor derived hPNF-MSCs maintained the fibroblast like morphology till late passage while deceased donor derived MSCs did not. Similarly growth kinetics was better in live donor derived hPNF-MSCs. Comparison of Immunophenotyping demonstrated consistent expression of MSC surface markers. Additional experiments strong expression of specific renal and islet cell markers in naïve hPNF-MSCs. Two conditions were adopted for renal differentiation (Adherent and spheroid cultures). cells were analyzed for early and late stage renal markers. hPNF- MSCs when with islet specific growth factors formed cell clusters staining positive for DTZ.

    • Taken together these results demonstrate deceased donor derived PNF a suggestive source of MSCs for specific research or therapeutic models. However certain modifications need to be implemented with the use of media components. To conclude based on the results obtained concomitant expression of renal and islet related proteins suggest their potential as a cellular therapy for treating Renal failure and diabetes- rising pandemic.

    Human Perinephric fat (hPNF-MSCs) - Novel source of mesenchymal stromal cells complemented with neuro-glial predisposition.
    • It is believed that each organ of the adult human body has its own niche of stem cells termed as mesenchymal stem/stromal cells. Interestingly, each source in many ways is inherently different from the others. These source and donor dependent biological differences are with a purpose and have given rise to the concept of tailor made research and therapeutic applications.

    • With this background, we attempted to isolate, characterize and differentiate mesenchymal stromal cells from a novel source, namely human perinephric fat, in terms of morphology, , karyotype, differentiation potential, senescence, and secretome profiling. To the best of our knowledge, this is the first complete characterization report of mesenchymal stromal cells from human perinephric fat. hPNF-MSCs were isolated from healthy screened with informed consent and propagated in vitro for a long term. Cells showed typical spindle shaped fibroblast like morphology, and tri-lineage differentiation potential till mid-late passage with persistent expression of surface markers - CD 44, CD 73, CD 90, CD 105, and CD 166 throughout cultivation, fulfilling the ISCT criteria for Mesenchymal stromal cells. Secretome analysis demonstrated secretion of pro-inflammatory and anti-inflammatory cytokines, chemokines, and growth factors. Additional experiments revealed strong expression of specific neuroglial markers in naïve hPNF-MSCs. Cells generated spheroids when plated at high density using the hanging drop method and remain viable in vitro, providing a useful research tool having suitable applications in drug discovery. Taken together, these results demonstrate a novel source suggestive of certain specific research/therapeutic models. Concomitant with the other groups, it is important to note that heterogeneity amongst donors exists and hence it is imperative to screen each sample for future applications. To conclude, concomitant expression of related proteins, supplemented with release of tropic factors of great importance by hPNF-MSCs, makes them a valuable asset in cellular therapy for range of neurodegenerative disorders.

    A comparative study of efficacy of wound healing by mesenchymal stem cell conditioned medium derived from bone marrow, Wharton's Jelly and Lipoaspirate in diabetic Wistar rats
    • Clinical application of mesenchymal stem cells is gaining faster momentum day by day. For the therapy of the unmet degenerative conditions of body to cosmetic applications, stem cells are finding its place. This novel therapy offers cure for numerous clinical conditions including wound healing. Even after the discovery of sizable number of exogenous agents for wound healing, their effective use in chronic, complicated wounds like diabetic wounds is still questionable. This made scientist to think about the alternatives like application of mesenchymal stem cells. Majority of studies in this field supports the theory of homing of stem cells to the site of injury and further action of stem cells mediated through its trophic factors. In invitro conditions these factors can be obtained from the cell culture media, the conditioned medium and this medium is now gaining importance as a very effective tool for regeneration. With the emergence of this approach we conducted a study which evaluated the comparative efficacy of mesenchymal stem cell conditioned medium from three different sources namely Bone marrow, Wharton’s Jelly and Lipoaspirate in a diabetic wound model in rats. We have performed the secretome profiling of the cell types used in the study and could demonstrate the presence of trophic factors which plays crucial role in different stages of wound healing. The test items were infused subcutaneously on the wound borders in specified frequency. We selected the morphometric analysis of wound area contraction and hisptopathological evaluation of wound post treatment as parameters for healing. Of the test items evaluated in the current study, conditioned medium obtained from Wharton’s jelly derived mesenchymal stem cells offers the most promising wound healing effect.

    • To conclude, we are reporting a one of its kind novel study which evaluated the comparative efficacy of mesenchymal stem cell conditioned medium from three different sources namely Bone marrow, Wharton’s Jelly and Lipoaspirate in a diabetic wound model in rats. Our finding of conditioned medium obtained from Wharton’s jelly derived mesenchymal stem cells as the most promising candidate of diabetic wound healing demands further studies which will critically evaluate the regenerative potential of Wharton’s Jelly mesenchymal stem cells and its conditioned medium.