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。
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.
出处
《生物技术通报》
CAS
CSCD
北大核心
2013年第7期94-98,共5页
Biotechnology Bulletin
基金
"重大新药创制"科技重大专项课题(2012ZX09301003-001-005)
关键词
鸡
胚胎干细胞
胚盘细胞
体外培养
鉴定
Chicken
Embryonic stem cells
Blastodermal cells
In vitro culture
Characterization
