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Cell Journal، جلد ۱۶، شماره Suppl ۱، صفحات ۱۵۸-۱۵۸
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عنوان انگلیسی Ps-127: Differentiation of Human Adipose Derived Mesenchymal Stem Cells to Insulin-Producing Cells
چکیده انگلیسی مقاله Objective: Diabetes mellitus type-1 or insulin-dependent diabetes is an autoimmune disease in which the beta cells of pancreas are attacked by the immune system and destroyed. To this date, no definitive way to cure the disease has been discovered. However, studies on the use of differentiated mesenchymal stem cells in the treatment of some diseases such as diabetes mellitus have emphasized. Evaluation of insulin-producing cell (IPCs) differentiation from human adipose mesenchymal stem cells (h-ASCs) in RNA and protein level is the main objective of this study. Materials and Methods: Adipose tissue was obtained by liposuction. After enzymatic digestion and isolation of mononuclear cells, characterization of these cells was performed with differentiation assay to adipocytes and osteocytes and also cell surface specific antigens expression was detected by flowcytometry. Differentiation process was created in two stages. Low glucose-DMEM supplemented with 20% FBS, betamercaptoethanol (1 mmol/L) and nicotine amide (10 mmol/ L) was used in pre-conditioning stage for 48 hours. Then, the final stage includes high glucose- DMEM with nicotine amide (10 mmol/L) without FBS was performed up to 28th day. For expression assay of specific gene, RT-PCR and gel electrophoresis were done. Furthermore, dithizone (DTZ) staining and ELISA test for insulin secretion assay were performed. Results: According to gel electrophoresis, expression of PAX4, Glut2, PDX-1, and insulin genes in mRNA level was observed. Bata-actin was performed as an internal control. ELISA data showed significant differences in insulin secretion at day 28 compared to day 14 as well as first day. DTZ-stained cellular clusters appeared after 28 days in the culture flask. Conclusion: Cellular and molecular experiments for differentiation of h-Ad-MSCs to IPCs were evaluated in this study. Finally, based on the obtained results it can be suggested that this method is appropriate and functional for IPCs differentiation from h-ASCs.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/452
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