Pdx1基因修饰的人脐带间充质干细胞体外诱导分化为胰岛β样细胞  

Induced Differentiation of Human Umbilical Cord Mesenchymal Stem Modified by Cells Pdx1Gene into Islet β-like Cells in Vitro

作　　者：王娟[1] 高杨军[1] 陆琰[1] 唐小龙[1] 何冬梅[1] 张洹[1] 

机构地区：[1]暨南大学医学院血液病研究所,广州510632

出　　处：《生物医学工程学杂志》2011年第6期1175-1180,1206,共7页Journal of Biomedical Engineering

基　　金：国务院侨办重点学科建设基金资助项目(51205002)

摘　　要：本研究探讨胰十二指肠同源框因子-1(Pdx1)基因修饰的人脐带间充质干细胞(MSCs)体外分化为胰岛β样细胞。采用携带Pdx1基因的重组腺病毒感染MSCs 7d,联合细胞因子分阶段诱导分化。RT-PCR及Westernblot、免疫细胞化学法检测诱导后细胞胰岛素相关基因表达变化;化学发光法检测诱导后细胞培养液上清中胰岛素和C肽的分泌水平。用流式细胞术检测胰岛素阳性细胞百分率。结果显示:诱导转入Pdx1基因的人脐带MSCs后,梭形细胞聚集形成胰岛样细胞团,双硫腙染色胞浆呈亮红色;诱导后的细胞表达Pdx1、胰岛素、C肽和葡萄糖转运体2基因;诱导细胞分泌胰岛素和C肽,并且在高糖作用后,胰岛素和C肽分泌增加;诱导后胰岛素阳性细胞百分率为(11.61±4.83)%。结果表明,Pdx1修饰人脐带MSCs在体外能够诱导分化为胰岛β样细胞。This study was to explore the induced differentiation of human mesenchymal stem cells （MSCs） modified by pancreatic and duodenal homeobox factor 1 （Pdxl） gene into insulin-producing cells in vitro. After recombined adenovirus vector with Pdxl gene infected MSCs for 7d, cells were induced by induction factors. The genes＇ expressions related to islet β cells such as Pdxl, 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 reeombined adenovirus with Pdxl 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 β cells, such as Pdxl, 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 Pdxl gene modified MSCs from human umbilical cord could be induced to differentiate into islet β-1ike cells.

关 键 词：人脐带 间充质干细胞 胰腺十二指肠同源框因子-1 胰岛β样细胞 腺病毒载体 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]