鸡胚胎干细胞的体外培养和鉴定

生物技术通报 ›› 2013, Vol. 0 ›› Issue (7): 94-98.

鸡胚胎干细胞的体外培养和鉴定

何文俊^1，2 刘红¹ 叶玲玲¹ 李世崇¹ 王启伟¹ 陈昭烈¹

（1.军事医学科学院生物工程研究所细胞工程研究室，北京 100071；2.解放军总医院第一附属医院创伤外科研究室，北京 100048）

收稿日期:2013-01-22 修回日期:2013-07-19 出版日期:2013-07-19 发布日期:2013-09-02
作者简介:何文俊，男，博士，研究方向：干细胞与再生医学；E-mail：520hewenjun@163.com
基金资助:
“重大新药创制”科技重大专项课题（2012ZX09301003-001-005）

Culture and Characterization of Chicken Embryonic Stem Cells in vitro

He Wenjun^1，2 Liu Hong¹ Ye Lingling¹ Li Shichong¹ Wang Qiwei¹ Chen Zhaolie¹

（1. Department of Cell Engineering，Institute of Biotechnology，Academy of Military Medical Sciences，Beijing 100071；2. Wound Healing and Cell Biology Laboratory，the First Affiliated Hospital，Chinese PLA General Hospital，Beijing 100048）

Received:2013-01-22 Revised:2013-07-19 Published:2013-07-19 Online:2013-09-02

摘要/Abstract

摘要： 从鸡X期胚胎中分离胚盘细胞，以小鼠胚胎成纤维细胞（mouse embryonic fibroblasts，MEF）为滋养层，高糖DMEM为基础培养基并添加10 ng/mL bFGF、20 ng/mL hIGF-1、2 ng/mL mSCF、2 ng/mL hIL-11和1 000 U/mL LIF等细胞因子对细胞进行培养，获得典型的呈集落生长的鸡胚胎干细胞（chicken embryonic stem cells，cESCs）克隆。免疫组化显示，呈集落生长的cESCs克隆具有较高的内源性AKP活性并表达多能性标志SSEA-1和Oct-4。RT-PCR分析进一步证实其表达cESCs特异性基因cENS-1。结果表明，该培养体系可用于cESCs的分离并维持其在体外生长并保持未分化状态。

关键词: 鸡, 胚胎干细胞, 胚盘细胞, 体外培养, 鉴定

Abstract: The blastoderm cells isolated from the stage X embryos of chicken were cultured on feeder layer of mouse embryonic fibroblasts（MEF）with high glucose DMEM supplemented with 10 ng/mL bFGF, 20 ng/mL hIGF-1, 2 ng/mL mSCF, 2 ng/mL hIL-11 and 1 000 U/mL LIF. Typical chicken embryonic stem cells（cESCs）colonies with high endogenous AKP activity were observed in the culture system. The cESCs derived from blastoderm cells expressed pluripotent markers SSEA-1 and Oct-4 as determined by immunocytological analysis. RT-PCR analysis further confirmed the expression of cENS-1, a gene specifically expressed in cESCs and early embryo. These results demonstrate the feasibility for using the culture system in isolating cESCs from the stage X chicken embryos and supporting the growth of cESCs with undifferentiating state in vitro.

Key words: Chicken, Embryonic stem cells, Blastodermal cells, In vitro culture, Characterization

何文俊,刘红,叶玲玲,李世崇,王启伟,陈昭烈. 鸡胚胎干细胞的体外培养和鉴定[J]. 生物技术通报, 2013, 0(7): 94-98.

He Wenjun,Liu Hong,Ye Lingling,Li Shichong,Wang Qiwei,Chen Zhaolie. Culture and Characterization of Chicken Embryonic Stem Cells in vitro[J]. Biotechnology Bulletin, 2013, 0(7): 94-98.

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