关键词: 关键词：血液细胞, 诱导多能干细胞, 胰岛素分泌细胞, Ⅰ型糖尿病

Abstract: Objective To generate and identify iPSCs from T1DM patient’s blood cells and to induce it into insulin-producing cells (IPCs) to treat the T1DM mice. Methods Erythroid progenitor cells from T1DM patient were infected by episomal vectors with an oriP/Epstein-Barr nuclear antigen-1 (oriP/Epstein-Barr nuclear antigen-1, oriP/EBNA-1) backbone for delivering the reprogramming genes Oct4, Sox2, Lin28, L-Myc and Klf4 to generate iPSCs. The multipotency of the iPSCs were verified by karyotyping analysis, alkaline phosphatase staining, RT-PCR and teratoma test. It was induced to differentiate into IPCs by 4 stages. To identify the IPCs by determining the marker genes expression of pancreatic β cell and Glucose-stimulated C-peptide release test. The IPCs were transplanted into the capsul of left kidney in the male C57BL/6J diabetic mouse model. Blood glucose and weight were continuously monitired. The glucose tolerance test was performed. Results The obtained iPSCs carried normal karyotype and expressed the pluripotent genes Oct4, Sox2, Lin28, L-Myc and Klf4. Alkaline phosphatase were positive. In the teratoma test, three germ layer cells were found. The obtained IPCs expressed the marker genes Pdx-1, Insulin and Nkx6.1. The glucose stimulated induced the secretion of C-peptide. The blood glucose was decreased and the weight was increasd after transplantation to the diabetic mice. The ability of regulating glucose was improved. Conclusion IPSCs form T1DM patient’s blood cells may induce into IPCs. It has a therapeutic effect after transplanting to the T1DM mice.

Key words: Key words：blood cells, induced pluripotent stem cells, insulin-producing cells, type 1 diabetes