1690-P: In Vitro Differentiation of Pancreas-Derived Mesenchymal Stem Cells into Insulin-Producing Beta Cells

Autor:	Prathab Balaji Saravanan, Jagan Kalivarathan, Mazhar A. Kanak, Marlon F. Levy
Rok vydání:	2020
Předmět:	geography geography.geographical_feature_category medicine.diagnostic_test Endocrinology Diabetes and Metabolism Mesenchymal stem cell CD34 Biology Islet Flow cytometry Cell biology medicine.anatomical_structure Adipogenesis Internal Medicine medicine CD90 Stem cell Pancreas
Zdroj:	Diabetes. 69
ISSN:	1939-327X 0012-1797
Popis:	Differentiation of insulin-producing cells (IPCs) from stem cells has been widely studied as an alternative source for cell transplantation therapy in diabetes. Mesenchymal stem cells (MSCs) are a useful therapeutic option for islet transplants due to their immunomodulatory properties and differentiation potential. In this study, we introduce a method for in vitro differentiation of pancreatic MSCs into IPCs. Research-grade pancreas from cadaveric donors was used to isolate islets, and the COBE remnant after islet purification was used for the generation of pancreatic MSCs. The remnant tissue was cultured in RPMI medium and plastic to isolate tissue-resident MSCs. Surface marker expression analysis and differentiation assays were performed to characteristically confirm that the cells obtained were in fact MSCs. Osteogenic, adipogenic and chondrogenic differentiation of MSCs were induced by StemPro® differentiation kits. The protocol for the differentiation of IPCs from stem cells was adopted from Pagliuca et al. 2014. Similarly, we have treated the pancreatic MSCs with molecules and growth factors at different stages of development to produce IPCs. The insulin secretion from differentiated IPCs also analyzed at all stages. The culture of remnant tissue after islet isolation resulted in plastic-adherent cells of fibroblast-like morphology after 3 passages. Flow cytometry analysis showed homogeneous expression of markers (CD90, CD73, CD29, CD105) and negative expression of CD14, CD34, and CD45 as classically characterized by cultured MSCs. Interestingly, there was a significant increase in insulin levels up to 5 and 100 folds in the media at developmental stages S5 (P= 0.0686) and S6 (P Disclosure J. Kalivarathan: None. P. Saravanan: None. M.F. Levy: None. M.A. Kanak: None.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::386c6afa269dc0ed411326670302f498 https://doi.org/10.2337/db20-1690-p Zobrazit plný text záznamu