重编程诱导犬脂肪间充质干细胞向胰岛素分泌细胞分化OA北大核心CSTPCD

The purpose of this study is to use transgenic technology to induce Adipose-derived mesenchymal stem cells(AMSCs)to differentiate into insulin secured cells(IPCs),and to achieve the purpose of clinical treatment of diabetes through stem cell therapy.This study first verified the role of Hnf1b gene in inducing the differentiation of canine AMSCs into IPCs in vitro,and on this basis combined with the cascade regulatory factors Pdx1,Ngn3,Pax4 and Nkx6.1,which play a key role in the development of islet cells,three different polygenic co-expressing adenovirus infected canine AMSCs.It was reprogrammed to differentiate into IPCs,and the optimal induction combination was selected by detection.The results showed that Hnf1b gene could promote the differentiation of canine AMSCs into IPCs in vitro.Three combinations of multi-gene co-expression vectors were constructed,pAdEasy-H nf1b-Pdx1-Ngn3(A),pAdEasy-Hnf1b-Pdx1-Ngn3-Pax4(B),pAdEasy-Hnf1b-Pdx1-Ngn3-Nkx6.1(C).After 25 days infected with canine AMSCs,pancreatic islet-like cell clusters were formed in group A,B,and C.Group B had the largest number,and dithizone staining was the deepest.The amount of insulin secretion in group B under high glucose stimulation was significantly higher than group A and group C.The expressions of genes related to islet cell development in groups A,B and C were significantly increased.The expression of Pdxl gene in group B was significantly higher than group A and group C;The Pcsk1 gene expression in group B was significantly higher than that in group C;The expression of Ins gene in group B was higher than that in group A.In summary,Hnf1b gene can promote the differentiation of canine AMSCs into IPCs in vitro.It is shown that the combination of Hnf1b,Pdx1,Ngn3 and Pax4 genes has the best induction efficiency,which provides a new reference for exploring more efficient methods of stem cell differentiation into IPCs,and provides new ideas for clinical treatment of diabetes.