Characterization of synovium-derived mesenchymal stem cells from joints with osteochondritis dissecans

Introduction: Osteochondritis dissecans (OCD) is a pathologic condition that occurs in children as well as adults. OCD is often managed based on the extent of ischemia and the stage of the disease. Synovial tissue collected during an arthroscopic procedure might serve as an ideal source for autologous mesenchymal stem cells (MSCs) with a potential for regenerative medicine of cartilage. Therefore, the present in vitro study aimed to evaluate the potency characteristics of synovium-derived MSCs (SMSCs) from joints with OCD for prospective autologous therapy. Methods: Primary culture of SMSCs was established and basic cellular properties, such as morphology, growth kinetics and clonal propagation ability, were analyzed. The expression of phenotypic markers, including CD29, CD44, CD90, CD34 and CD45, was assessed by flow cytometry and immunocytochemistry. Mesodermal differentiation into osteocytes, chondrocytes and adipocytes was performed using standard protocols. Expression of chondrocyte-specific markers was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Isolated SMSCs displayed fibroblast-like morphology with > 95% cell viability and had high proliferative rates with a shorter doubling time. The cells showed positive expression of CD29, CD44 and CD90, but were negative for CD34 and CD45 markers. Upon induction, SMSCs were successfully differentiated into osteogenic, chondrogenic and adipogenic lineages. Chondrogenesis was more prominent in SMSCs than osteogenesis. Chondrogenesis was further confirmed by the expression of aggrecan and collagen type IIα1 markers. Conclusion: SMSCs showed greater proliferation and an enhanced ability for chondrogenic differentiation. Synovium can be harvested with minimal tissue damage and donor-site morbidity and might serve as an alternative autologous source of MSCs for cartilage-tissue regeneration.

Systemic Administration of Rejuvenated Adipose-Derived Mesenchymal Stem Cells Improves Liver Metabolism in Equine Metabolic Syndrome (EMS)- New Approach in Veterinary Regenerative Medicine

Equine metabolic syndrome (EMS) is characterized by adiposity, insulin dysregulation and increased risk for laminitis. Increased levels of specific liver enzymes in the peripheral blood are typical findings in horses diagnosed with EMS. Current management of EMS is based on caloric restriction and increased physical activity. However, new potential treatment options are arising such as the transplantation of autologous adipose stem cells (ASC). However, cytophysiological properties of ASC derived from EMS horses are impaired which strongly limits their therapeutic potential. We hypothesized, that in vitro pharmacotherapy of those cells with 5-azacytidine (AZA) and resveratrol (RES) before their clinical application can reverse the aged phenotype of those cells and improve clinical outcome of autologous therapy. A 9 year old Dutch Warmblood Horse used for driving, was presented with severe obesity, insulin resistance. After EMS diagnosis, the animal received three intravenous injections of autologous, AZA/RES treated ASCs at weekly intervals. The therapeutic effect was assessed by the analysis of liver specific enzymes in the blood. ASC-transplantation reduced levels of glutamate dehydrogenase (GLDH), gamma-glutamyltransferase (GGT), lactate dehydrogenase (LDH) and aspartate transaminase (AST). This case report demonstrates the therapeutic potential of this intervention for EMS as well as apt utility of autologous, rejuvenated ASC injections.

Comparison of proliferative and immunomodulatory potential of adipose-derived mesenchymal stem cells from young and geriatric cats

Objectives The objective of this study was to compare the ability of adipose-derived mesenchymal stem cells (aMSCs) generated from young vs geriatric cats to proliferate in culture, suppress lymphocyte proliferation and undergo senescence. Methods Adipose tissues from five young (<5 years) and six geriatric (>10 years) cats were harvested and cryopreserved for subsequent aMSC isolation and culture. aMSC proliferation in culture was compared via determination of time until passage two and by 3-(4,5-demethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The immunomodulatory capacity of aMSCs was assessed using lymphocyte proliferation assays, and senescence was evaluated using senescence-associated B-galactosidase (SABG) expression. All assays were performed on aMSCs between passage two and passage three. Results aMSCs from geriatric cats took significantly longer ( P = 0.008) to reach passage two (median 11 days, range 9–22 days) compared with aMSCs from young healthy cats (median 7 days, range 6–8 days). No significant difference was detected between young and geriatric cats in terms of their ability to suppress lymphocyte proliferation. SABG expression was not significantly different between young and geriatric aMSCs. Conclusions and relevance Compared with young feline aMSCs, geriatric aMSCs are significantly impaired in their ability to rapidly proliferate to passage two following initial culture, presenting a concern for autologous therapy. Nonetheless, once the cells are expanded, young and geriatric cat aMSCs appear to be equivalent in terms of their ability to functionally suppress T-cell activation and proliferation.