Abstract:

Objective To isolate human adipose tissue-derived stromal cells (hADSCs) from the adipose tissue in vitro,and to induce the differentiation of hADSCs into insulin-producing cells,and to study the potential of hADSCs to differentiate into insulin-producing cells.Methods hADSCs were isolated from the adipose tissue.The hADSCs were differentiated to islet-like cells using basic fibroblast growth factor(bFGF),epidermal growth factor (EGF),nicotinamide and exendin-4.RT-PCR analysis was used to detect the expressions of genes in differentiation,such as pancreatic and duodenal homeobox 1(Pdx-1),insulin and glucose transporter 2(Glut-2);the differentiated cells were tested by Dithizone staining;Immunocytochemical analysis was performed to determine the insulin and Pdx-1;ELISA was done to assay the insulin secretion in response to glucose stimulation.Results hADSCs became polygon from fibroblast-like about 7 d after treated with the induced medium,and began to gather and started to form cell aggregates,the islet-like cell clusters were formed at 21 d.Dithizone staining was positive;RT-PCR analysis showed the expressions of the insulin,Pdx-1,Glut-2 and glucagon in the differentiated cells.Immunocytochemical analysis showed that the induced cells expressed insulin and Pdx-1.These cells secreted insulin in response to glucose stimulation,and the levels of insulin secretion were increased with the increasing of glucose concentrations.Conclusion hADSCs isolated from adipose tissues can be induced to differentiate into insulin-producing cells,and they can be used as a potential source for transplantation in the patients with diabetes mellitus.

Key words: adipose tissue-derived stromal cells;insulin-producing cells;differentiation