1. 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.
- Author
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Gupte KS, Vanikar AV, Trivedi HL, Patel CN, and Patel JV
- Subjects
- Adipose Tissue immunology, Cell- and Tissue-Based Therapy, Humans, Immunomodulation immunology, Mesenchymal Stem Cells immunology, Adipose Tissue cytology, B-Lymphocytes immunology, Interleukin-10 metabolism, Leukocytes, Mononuclear immunology, Mesenchymal Stem Cells cytology
- Abstract
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., (Copyright © 2017 Chang Gung University. Published by Elsevier B.V. All rights reserved.)- Published
- 2017
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