In-vitro generation of interleukin-10 secreting B-regulatory cells from donor adipose tissue derived mesenchymal stem cells and recipient peripheral blood mononuclear cells for potential cell therapy.

BACKGROUND: Interleukin-10 secreting B-cells are a major subset of B-regulatory cells (B-regs), commonly recognized as CD19+/38hi/24hi/IL10+. They carry out immunomodulation by release of specific cytokines and/or cell-to-cell contact. We have generated B-regs in-vitro from donor adipose tissue derived mesenchymal stem cells (AD-MSC) and renal allograft recipient (RAR) peripheral blood mononuclear cells (PBMC) for potential cell therapy. MATERIAL AND METHODS: Mononuclear cells separated by density gradient centrifugation from 50 ml anti-coagulated blood of 15-RAR and respective donors were analysed for baseline B-regs using appropriate antibodies. Equal amount (20 × 106 cells/ml) of stimulator (irradiated at 7.45 Gy/min for 10 min) and responder (non-irradiated) cells were co-cultured with in-vitro generated AD-MSC (1 × 106 cells/ml) in proliferation medium containing lipopolysaccharide from E. coli K12 strain at 37 °C with 5% CO2. Cells were harvested on day-7 and analyzed for viability, sterility, quantity, morphology and phenotyping. In-vitro generated B-reg levels were compared with baseline B-regs. RESULTS: In-vitro generated B-reg count increased to 16.75% from baseline count of 3.35%. CONCLUSION: B-regs can be successfully generated in-vitro from donor AD-MSC and RAR PBMC for potential cell therapy.

In vitro Generated Mesenchymal Stem Cells: Suitable Tools to Target Insulin Dependent Diabetes Mellitus?

A synergy of a pre-accumulated genes with an autoimmunity advancing to slow abolition of pancreatic beta-cells causes insulin deficiency and results enrooting insulin dependent diabetes mellitus (IDDM). As per WHO data worldwide about 150 million people are diabetic and the number may rise to more than double by the year 2025. Any absolute cure for IDDM is not available yet, and one of the credible advent in the field include cell-based therapy. At this conjecture, mesenchymal stem cells (MSC) seems to have a specific and beneficial characteristics due to their in vivo as well as in vitro potential to mimic a pancreatic endocrine phenotype and immune-regulatory actions. MSC have the capacity to tweak endogenous tissue and cells of immune system. They have been proven as secure and efficacious cell-based regenerative therapy, to treat diverse autoimmune, degenerative diseases and tissue injuries. By consolidating characteristics of MSC biology, MSC-based therapy, engineering and advances in the field, MSC have a great potential to bring us notably closer to a much-needed and long-time awaited cure of IDDM. The review discusses MSC-based cellular therapeutic strategies targeting at IDDM. MSC characteristics of immunomodulation and regeneration potential when used alone or in combination with islets or in differentiated form of insulin producing cells (IPC) are taken into consideration for the review purpose.