鸡胚胎干细胞的分离培养与鉴定  

Isolation,culture and identification of chicken embryonic stem cells

作　　者：李欣 于永生[1] 吕阳 柳俭强 金海国[1] 朴庆林[1] 赵中利[1] LI Xin;YU Yong-sheng;LYU Yang;LIU Jian-qiang;JIN Hai-guo;PIAO Qing-lin;ZHAO Zhong-li(Branch of Animal Husbandry,Jilin Academy of Agricultural Sciences,Gongzhuling City,Jilin 136100,China;College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]吉林省农业科学院畜牧科学分院,吉林公主岭市136100 [2]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《中国兽医学报》2020年第6期1228-1231,共4页Chinese Journal of Veterinary Science

基　　金：吉林省农业科学院创新工程资助项目(CXGC-2018DC001)。

摘　　要：利用胰蛋白酶消化法从X期鸡胚中分离胚胎干细胞(embryonic stem cells,ESCs),以鸡胚成纤维细胞为滋养层,进行体外培养,采用形态法和标志分子检测对获得的干细胞进行鉴定。结果显示,获得典型的呈巢状或岛状的鸡胚胎干细胞(chicken embryonic stem cells,cESCs)克隆,细胞碱性磷酸酶(alkaline phosphatase,AKP)染色呈蓝紫色,表明具有较高的内源性AKP活性;胚胎阶段特异性表面抗原SSEA-1(stage specific embryonic antigen 1,SSEA-1)鉴定结果呈阳性,显示cESCs克隆具有多能性。本试验成功分离出cESCs,可体外培养并保持未分化状态及多能性。Embryonic stem cells(ESCs)were isolated from stage X chicken embryo by trypsin digestion,and cultured in vitro with chicken embryo fibroblasts as trophoblast,and the stem cells obtained were identified by morphological method and marker molecular detection.The results showed that typical nests or islands of chicken embryonic stem cells were obtained,and the cell alkaline phosphatase(AKP)stain was bluish-purple,indicating a high endogenous AKP activity.The result of embryonic SSEA-1 is positive,showing that the cloning of cESCs is pluripotent.This experiment successfully isolated chicken embryonic stem cells,which can be cultured in vitro and remain undifferentiated and pluripotent.

关 键 词：鸡 胚胎干细胞 体外培养 碱性磷酸酶 胚胎阶段特异性表面抗原 

分 类 号：S831.2[农业科学—畜牧学]