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Abstract • MSCs were isolated from bone marrow of 3 nondiabetic donors and cultured until reaching nearly 70% confluence. At the third passage, cultured cells were phenotypically characterized by flow cytometry using antibodies against CD105, CD45, and CD34. Then, we allocated cells into control group that were cultured in complete DMEM and differentiation group that were cultured in differentiation media containing several growth factors according to the three-stage differentiation protocol that lasted for 18 days. • During the stages of differentiation, the cultured cells clustered gradually in groups to form cellular aggregates. At the end of the differentiation protocol (18 days), the cells attained a spheroidal morphology, while control remain fibroblastic-shaped. • At the end of each stage of differentiation, the cells were harvested and total RNA was extracted to investigate the stepwise relative gene expression of PDX1, PAX4, and Insulin genes. At the end of differentiation period, differentiated cells were evaluated for C-peptide release under glucose stimulation. |