鸡胚培养与嵌合体鸡的制备  

Studies on Culture of Chick Embryos and Production of Chimeric Chickens

作　　者：贾彦征 熊一力 刘光泽 王洪敏 

机构地区：[1]空军广州医院全军传染病中心转基因工程研究室,广州510602

出　　处：《中国实验动物学杂志》2001年第2期93-94,共2页Chinese Journal of Laboratory Animal Science

摘　　要：目的　探讨自X期鸡胚培养方法 ,制备嵌合体鸡。方法　分离供体麻鸡种蛋的胚盘细胞 ,注入受体来航鸡种蛋胚盘 ,然后用以下两种方法培养鸡胚 :①钝端开窗法 ,沿鸡蛋气室边缘开口 ,内壳膜开小孔 ,注射胚盘细胞后再封口 ,置孵化器内 ,温度 38℃ ,翻蛋角 90° ;②两次倒蛋法 ,胚盘细胞注入受体蛋后 ,倒入代用蛋壳Ⅰ ,置孵化器内 ,温度 38℃ ,翻蛋角 90° ,72h后 ,倒入代用蛋壳Ⅱ ,改翻蛋角 30°。结果　用钝端开窗法注射 83只种蛋 ,出雏 35只 (4 2 2 % ) ,表型嵌合体 7只 ,3只存活 ;用两次倒蛋法 ,注射 72只种蛋 ,出雏 10只 (13 9% ) ,表型嵌合体 5只 ,1只存活。结论　钝端开窗法较经典的两次倒蛋法出雏率明显提高 ,用该方法制备嵌合体鸡更为方便可行。Objective\ To study culture system for chick embryos and produce chimeric chickens.Methods\ Blastodermal cells were isolated from stage X embryos of Ma chickens and injected into embryos of Leghorns.Two culture methods were used.(1)blunt\|end windowed.1.5cm window was made in the blunt\|end of the eggshell,the inner membrane was ripped a 0.5cm hole,so that the blastoderm was exposed,and blastodemal cells were injected into the subgerminal cavity.The window was closed with wrap film,then the embryos were hatched at 38℃,90° egg\|turning.(2)Twice transfer of embryo.The embryo was transferred into the alternative eggshell Ⅰ,blastodermal cells were injected,hatched at 38℃,90° egg\|terning for 72 hours,then transferred into alternative eggshell Ⅱ,hatched at 30° egg\|turning.Results\ Using method Ⅰ,of 83 injected embryos,35(42.2%)had survived to hatching,7 were phenotypically chimeric with respect to feather color and 3 survived.Using method Ⅱ,of 72 injected embryos,10(13.9%) survived,5 were chimeric and one survived.Conclusions\ The method of blunt\|end windowed improved the rate of hatching significantly,compared with the ancient method of twice transfer.This method made the production of chimeric chickens easily.

关 键 词：种蛋 翻蛋 胚培养 出雏 代用蛋壳 胚盘 鸡种 嵌合体 开窗法 注射 

分 类 号：Q813[生物学—生物工程] S831[农业科学—畜牧学]