Cdyl基因敲除的小鼠诱导多能干细胞系的建立及鉴定  

Establishment and Characterization of Induced Pluripotent Stem Cell Line from Cdyl Knockout Mice

作　　者：胡晓俊[1] 赖文玉[2] 万利[1] 王涛[1] 张秀明[1] 李伟强[1] 项鹏[1] 

机构地区：[1]中山大学干细胞与组织工程中心,广东广州510080 [2]中山大学附属第二医院儿科,广东广州510120

出　　处：《中山大学学报（医学科学版）》2011年第2期146-154,共9页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金博士启动(9451008901002230);国家自然科学基金(81000150)

摘　　要：【目的】建立Cdyl基因敲除的小鼠诱导多能干细胞系(iPSC)。【方法】通过组织块贴壁法培养Cdyl-/-的胚胎成纤维细胞(MEF)。将pMx-Oct3/4、Sox2、c-Myc、Klf4和hrGFP五种质粒分别转染Plat-E细胞,收集病毒上清,感染Cdyl-/-MEF获得Cdyl-/-iPSC,并对Cdyl-/-iPS细胞的分子标记及体内外分化能力进行检测。【结果】取Cdyl-/-胎鼠皮肤,培养得到增殖旺盛的Cdyl-/-MEF。成功包装逆转录病毒并感染Cdyl-/-MEF,得到胚胎干细胞样的Cdyl-/-iPS细胞。该细胞表达AKP、Oct3/4、SSEA-1,在体内外可以分化为三胚层的细胞类型。【结论】Cdyl-/-iPS细胞具有自我更新和多向分化能力的特性。Cdyl-/-iPS细胞系的建立为研究Cdyl在小鼠早期胚胎发育中的功能提供一个良好的细胞模型。[Objective]This study was designed to establish induced pluripotent stem cell line from Cdyl knockout mice.[Methods] Cdyl-/-MEF were attained by adhesion culture of E18.5 embryonic tissue.pMx-Oct3/4,Sox2,c-Myc,Klf4,and hrGFP plasmid were transfected into Plat-E cells,respectively.Then virus-containing supernatants were harvested to infect Cdyl-/-MEF.The characteristics of Cdyl-/-iPSC were analyzed by detection of pluripotent marker expression and differentiation capacity in vivo and in vitro.[Results] The Cdyl-/-iPS cell colonies were emerged 5 days after Cdyl-/-MEF were infected with virus-containing supernatants.Cdyl-/-iPSC expressed AKP,Oct3/4,SSEA-1 and were able to differentiate into cell types of three germ layers.[Conclusion] Cdyl-/-iPS cell line was successfully established and provides a valuable model for studying Cdyl function in mouse embryonic development.

关 键 词：小鼠诱导多能干细胞 Cdyl 基因敲除 小鼠胚胎成纤维细胞 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]