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• W2616759822 abstract "Introduction: During the recent years, Mesenchymal Stem cell (MSC) has gained importance in tissue repair, tissue engineering and in immunosuppressive therapy. Autologous MSCs are being used in the treatment of various diseases, including cardiovascular diseases, acute renal failure, autoimmune diseases, neurodegenerative disorders and organ transplant. Recently, ex-vivo expanded MSC infusion in renal transplant cases is being explored as a possible intervention for induction phase. Past studies support the feasibility of lowering the dose or avoiding the use of immunosuppressive drugs after induction with MSCs. Several reports also demonstrate that MSCs lead to decreased acute rejection rates, better graft function and decreased infection rates. The main objective of the study was to isolate, expand and characterize BM-derived autologous MSCs from CKD (Chronic kidney disease) patients for clinical translation. Methods: Bone marrow aspirates of the patients collected aseptically from the posterior iliac crest. Samples were centrifuged and the cellular layer containing the MSCs was carefully removed, washed, counted and seeded in low glucose Dulbecco modified Eagle medium (DMEM) containing 10% serum albumin and 1%antibiotic-antimycotic agents. After three passages, quality assurance was done as per the International Society of Cell therapy and transplantation. The seeding density was ~5 X 106 cells in T75 flasks. MSCs were allowed to adhere to the surface of the flasks for 72 hours at 37°C and 5% CO2. The non-adherent cells in the media were removed, fresh media added, incubated till the cells became confluent. At 70-80% confluence, cells were detached and replated ~25,000 MSCs in T 75 flask. Results: Approximately 10X106 cells were obtained after three passages in 1 month’s time frame without any contamination. The viability of thawed cells was 85.5%, and they maintained the stemness even after 6 months of cryopreservation similar to fresh MSCs. They expressed CD44, CD73, and CD90 and were negative for CD19, CD34, CD45, and HLA DR. Cytogenetic analysis showed no chromosomal abnormalities. Conclusion: Optimization of the protocol for isolation, expansion of MSCs from bone marrow has been done successfully for implementing MSC-based therapy in kidney transplantation, and to promote its clinical translation.FIGURE 1: Microscopic Images of Mesenchymal Stem Cells.FIGURE 2: Enumerarion of MSC-specific CD markers (CD90, CD-73 and CD44) in cells from 3rd passage of patient No.10 in Panel B shows viablity of cells as stained by 7-AAD. Further Panel B and C shows MSC-negative markers for MSCs.FIGURE 3: Karyotype report showing no chromosomal abnormality." @default.
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• W2616759822 date "2017-05-01" @default.
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• W2616759822 title "Isolation, Characterization and Expansion of Bone-Marrow derived Mesenchymal Stem cells (MSCs) from Chronic Kidney Disease (CKD) patients for Clinical Translation" @default.
• W2616759822 doi "https://doi.org/10.1097/01.tp.0000520387.31474.f3" @default.
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