腺病毒转染MafA诱导人脐带间充质干细胞表达胰腺细胞特异性基因

Adenovirus transfection of MafA induces human umbilical cord mesenchymal stem cells to express pancreatic cell-specific genes

背景:人脐带间充质干细胞经化学药物或共培养等方法诱导后胰岛素分泌量低,转染胰腺发育特异性基因诱导干细胞向成熟β细胞分化是近年来的研究热点。目的:探讨MafA基因转染后人脐带间充质干细胞向胰岛素分泌细胞分化的潜能。方法:腺病毒携带外源性β细胞发育的关键转录因子Maf A(Ad-MafA-EGFP),转染入第3代人脐带间充质干细胞,诱导后7 d观察细胞的形态学变化,应用PCR检测细胞中胰腺细胞特异性基因(glucagon、Pdx1、Nkx2.2)的表达。结果与结论:(1)经Ad-Maf A-EGFP转染后,倒置相差显微镜下观察人脐带间充质干细胞形态未见明显变化;(2)荧光显微镜下观察Ad-MafA-EGFP已进入细胞内;(3)经Ad-MafA-EGFP诱导后,人脐带间充质干细胞中人源性胰腺前体细胞glucagon、Pdx1、Nkx2.2基因表达增高,为人脐带间充质干细胞进一步分化成熟胰腺β细胞提供实验依据。

BACKGROUND： Human umbilical cord mesenchymal stem cells（HUMSCs） can differentiate into insulin-producing cells after induced by chemical drugs or co-culture methods, but insulin secretion is extremely low. Therefore, to induce mature pancreatic beta cell differentiation from stem cells by adenovirus transfection of specific genes involved in the development of pancreas is a research hotspot in recent years.OBJECTIVE：To study the differentiation potential of HUMSCs into insulin-producing cells after transfection with musculoaponeurotic fibrosarcoma oncogene homologue A（Maf A）.METHODS：Ad-Maf A-EGFP was transferred into passage 3 HUMSCs.After 7 days of induction,changes of cell morphology were observed by inverted phase contrast microscope.Expression of pancreatic cell-specific genes（glucagon,PDX1,Nkx2.2）was detected by PCR technique.RESULTS AND CONCLUSION：After Ad-Maf A-EGFP transfection,no significant morphological changes were observed in the HUMSCs under inverted phase contrast microscope.It was confirmed by fluorescence microscope that Ad-Maf A-EGFP was transferred into the HUMSCs.After induction,the expression of human pancreatic precursor cell-related genes,including glucagon,PDX1 and Nkx2.2,was increased as detected by PCR.To conclude,these findings could provide experimental evidence for further differentiation and maturation of pancreaticβcells from HUMSCs.