4种全能性基因转入人胚胎成纤维细胞诱导多能性干细胞系的建立及其鉴定(英文)  

Generation and identification of pluripotent stem cells from human embryonic fibroblast cells by 4 defined factors

作　　者：杜丽丽[1] 林戈[1,2] 卢光琇[1,2] 

机构地区：[1]中南大学生殖与干细胞工程研究所,长沙410078 [2]人类干细胞国家工程研究中心,长沙410078

出　　处：《中南大学学报（医学版）》2009年第12期1157-1165,共9页Journal of Central South University ：Medical Science

基　　金：supported by the Ministry of Science and Technology of China(2006AA02A102);the National Basic Research Program of China(2005CB522705)

摘　　要：目的:利用人类胚胎成纤维细胞(human embryonic fibroblast cells,hEFs)获得诱导性多能干细胞(induced pluripotent stem cells,iPSCs)系并对其全能性进行鉴定。方法:本研究采用慢病毒感染的方法将含有Oct4,Sox2,Nanog和Lin28全能性基因的慢病毒颗粒转导hEFs,获得iPSCs,并对其进行碱性磷酸酶(alkaline phosphatase,AKP)染色,以及检测表面标记,端粒酶活性,EB分化和畸胎瘤形成等,同时进行核型分析以及短串联重复序列(short tandem repeat,STR)分析。结果:所获得的iPSCs表达多能性细胞的表面标记,具有高的端粒酶活性,可在体内体外向内中外三胚层分化,与hESCs相似。经STR分析证实,iPSCs确实来源于hEFs而非胚胎干细胞的污染。结论:4种全能性基因转入hEFs可诱导获得与胚胎干细胞相似的iPSCs,有助于进一步的表观遗传学重编程以及干细胞多能性维持的研究。Objective To generate and identify the induced pluripotent stem cells（iPSCs）from human embryonic fibroblast cells（hEFs）by introducing 4 defined factors.Methods We introduced 4 factors（Oct4,Sox2,Nanog,and Lin28）into hEFs by lentivirus infection.The iPSCs generated from this method were analyzed in many aspects,including surface antigens,gene expression,and telomerase activity,differentiation ability in vivo and in vitro and the patterns of short tandem repeat（STR）.Results The human iPSCs generated from this study were positive for alkaline phosphatase（AKP）staining,expressed hESCs-specific surface antigens,and exhibited high telomerase activity.They also possessed the ability to differentiate into 3-germ layers both in vivo and in vitro.STR analysis indicated that the human iPSCs were derived from the donor material.Conclusion The iPSCs lines can be obtained from human embryonic fibroblast by introducing 4 factors.Such human iPSCs should be useful in the study of epigenetic reprogramming and pluripotency maintenance.

关 键 词：诱导性多能干细胞 慢病毒感染 胚胎干细胞 胚胎成纤维细胞 

分 类 号：Q813[生物学—生物工程]