摘要
目的提高人诱导性多能干细胞(hiPSC)转化效率,缩短其产生周期,并避免引入异源基因。方法利用GP2-293反转录病毒包装系统对人包皮成纤维细胞(HFFs)进行转化,引入小分子物质,并对类胚胎干细胞(ESC)克隆进行独立培养,检测标记蛋白,定向诱导分化能力和核型分析。结果 HFFs经转化后,20 d出现形态与ESC克隆相似的hiPSC,比经典法缩短了10 d,效率提高约12倍。它们与ESC同样表达标记蛋白;体外成功定向分化为心肌细胞。核型分析表明该hiPSC染色体核型正常。结论建立稳定的转化效率高、周期短的hiPSC生成体系,并成功诱导分化成心肌细胞,为心血管疾病的模型构建和临床研究提供实验基础。
Objective To increase the transduction efficiency of hiPSC formation, decrease the time schedule and avoid the possibility of exogene introduction. Methods Human fibroblast cells were infected by retrovirus with four factors. During transduction, small molecules were added into hESC medium, until iPSC colonies grew big enough to be transferred. The stable iPSC colonies were detected by the expression of ESC protein markers, cardiomyocytes differentiation and karotyping analysis. Results The reprogramming period was shortened by 10 days with 12 times higher efficiency. The iPSC shared similar protein expression level with hESC. Both iPSC and hESC were differen- tiated into cardiomyocytes successfully with similar MEA data and both showed normal karotyping results. Conclu- sions The established iPSC reprogramming system is efficient and time-saving for providing research model for car- diovascular disease modeling and clinical study.
出处
《基础医学与临床》
CSCD
北大核心
2012年第9期1030-1035,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(81072510)
河南省教育厅课题(2008A340001)
关键词
诱导性多能干细胞
转化效率
形成周期
心肌细胞
分化
induced pluripotent stem cells
transduction efficiency
time schedule
cardiomyocytes
differentiation
