鸡胚胎干细胞的分离及其嵌合体制备条件探索  

Isolation of Chicken Embryonic Stem Cell and Exploring the Preparation Conditions of Chicken Chimeric

作　　者：张亚妮[1] 杨海燕[1] 施青青[1] 张振韬[1] 郑蒙蒙[1] 王丹[1] 李碧春[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《中国畜牧杂志》2012年第21期28-31,共4页Chinese Journal of Animal Science

基　　金：高等学校博士学科点专项科研基金资助课题(20103250110006);江苏省"六大人才高峰";中国博士后基金(2012M11806)

摘　　要：分离培养鸡胚胎干细胞(ESCs),体外培养传代后,进行碱性磷酸酶活性检测和SSEA-1染色鉴定;并用线性化的质粒pEGFP-N1转染鸡ESCs。受体种蛋经赤道面开窗后,将经转染的ESCs注射到受体鸡胚胚下腔,以便制作嵌合体鸡;对获得的嵌合体鸡分别提取血液和组织DNA后进行PCR检测。结果表明:5只存活的的嵌合体鸡血液中没有发生EGFP基因嵌合,但有4只嵌合体鸡的部分器官表达了EGFP基因,其中有2只鸡的性腺发生嵌合,表明利用赤道面开窗后对受体鸡进行胚下腔显微注射可以生产嵌合体鸡。Chicken embryonic stem cells （ESCs） were separated and cultured in vitro. Alkaline phosphatase activity （AKP） and SSEA-1 staining was conducted to detected ESCs. Then, chicken ESCs were transfected with linearized plasmid pEGFP-N1. The receptor eggs were drilled a window at the lateral shell of egg, then, the ESCs transfected with pEGFP-N1 was injected into chick embryo cavity so as to make chimeric chicken. The chimeric chicken was de- tected for both of blood and different tissue by PCR. The results showed five chimeric chicken were obtained in this study and found that four of them expressed EGFP gene in some organs, only two of them expressed EGFP gene in the gonad, indicating the chimeric chicken could be obtained through chick embryo cavity micro-injection by drilling a window at the lateral shell of egg.

关 键 词：鸡胚胎干细胞 分离培养 显微注射 嵌合体鸡 

分 类 号：S831.3[农业科学—畜牧学]