Wang Juan, Gao Yangjun, Lu Yan, Tang Xiaolong, He Dongmei, Zhang Yuan
Institute of Hematology, Medical College, Jinan University, Guangzhou 510632, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2011 Dec;28(6):1175-80, 1206.
This study was to explore the induced differentiation of human mesenchymal stem cells (MSCs) modified by pancreatic and duodenal homeobox factor 1 (Pdx1) gene into insulin-producing cells in vitro. After recombined adenovirus vector with Pdx1 gene infected MSCs for 7 d, cells were induced by induction factors. The genes' expressions related to islet beta cells such as Pdx1, insulin, glucose transporter-2 (Glut2), were detected with RT-PCR, immunocytochemistry and Western blot. The levels of insulin and C peptide secretion were examined with chemiluminescence immunoassay. Insulin(+) cell rate was detected by flow cytometry. After infected by recombined adenovirus with Pdx1 and combined with induction factors, MSCs were aggregated and islet-like cell clusters formed. Dithizone staining of these cells was positive. The genes' expression related to islet beta cells, such as Pdx1, insulin, Glut2, could be detected. After induction, the islet-like cell clusters secreted insulin and C peptide. The levels of insulin and C peptide secretion increased with glucose stimulation. Insulin(+) cell rate was (11.61 +/- 4.83)%. It could be concluded that Pdx1 gene modified MSCs from human umbilical cord could be induced to differentiate into islet beta-like cells.
本研究旨在探讨经胰腺十二指肠同源盒因子1（Pdx1）基因修饰的人骨髓间充质干细胞（MSCs）在体外诱导分化为胰岛素分泌细胞的情况。用携带Pdx1基因的重组腺病毒载体感染MSCs 7天后，用诱导因子进行诱导。采用逆转录聚合酶链反应（RT-PCR）、免疫细胞化学和蛋白质免疫印迹法检测与胰岛β细胞相关的基因如Pdx1、胰岛素、葡萄糖转运蛋白2（Glut2）的表达。采用化学发光免疫分析法检测胰岛素和C肽的分泌水平。通过流式细胞术检测胰岛素（+）细胞率。用携带Pdx1的重组腺病毒感染并结合诱导因子后，MSCs聚集形成胰岛样细胞团。这些细胞的双硫腙染色呈阳性。可检测到与胰岛β细胞相关的基因如Pdx1、胰岛素、Glut2的表达。诱导后，胰岛样细胞团分泌胰岛素和C肽。胰岛素和C肽的分泌水平随葡萄糖刺激而升高。胰岛素（+）细胞率为（11.61±4.83）%。可以得出结论，Pdx1基因修饰的人脐带间充质干细胞可被诱导分化为胰岛β样细胞。
