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该文通过Tet调控下共表达PDX1与BTC的骨髓间充质干细胞系(PDX1~+BTC~+MSCs),探讨PDX1和BTC共表达对骨髓间充质干细胞分化为胰岛素分泌细胞(IPCs)的效率及成熟度的影响。采用两步法对PDX1~+BTC~+MSCs细胞系诱导分化成IPCs,第一步Dox诱导7天检测到Nestin、CK19表达;第二步再诱导7天后形成DTZ染色阳性的胰岛样结构,Ngn3、Nkx6.1 mRNA水平和PDX1、Insulin、Glucagon的蛋白表达阳性。分化后的IPCs在葡萄糖刺激下能产生胰岛素和C肽,但仍不能达到正常胰岛水平。提示利用Tet-On体系调控PDX1和BTC共表达对骨髓间充质干细胞进行修饰,能有效诱导骨髓间充质干细胞分化为胰岛素分泌细胞,但分化成熟度仍然与天然胰岛细胞功能存在差距。
In this study, PDX1 ~ + BTC ~ + MSCs co-expressing PDX1 and BTC cells under Tet regulation were used to explore the effect of co-expression of PDX1 and BTC on differentiation of bone marrow mesenchymal stem cells into insulin-secreting cells (IPCs) And the maturity of the impact. The two-step method was used to induce the differentiation of PDX1 ~ + BTC ~ + MSCs into IPCs. The first step was to detect the expression of Nestin and CK19 in Dox for 7 days. The second step was to induce DTZ-positive islet-like structure , Nkx6.1 mRNA level and protein expression of PDX1, Insulin and Glucagon were positive. Differentiated IPCs produce insulin and C-peptide upon glucose stimulation but still fail to reach normal islet levels. It is suggested that the regulation of PDX1 and BTC co-expression by Tet-On system can modify BMSCs and induce bone marrow mesenchymal stem cells to differentiate into insulin-secreting cells. However, the differentiated maturation still has a gap with the function of native islet cells.