牛与小鼠胚胎在分化抑制培养系统中的行为比较

摘要

在自制培养基中，以原代小鼠胎儿成纤维细胞为饲养层，观察比较了牛与小鼠胚胎在体外分化抑制培养体系中的生长行为。结果表明，牛囊胚一般培养36～60 h后孵化脱带，96～120 h后贴壁，120～144 h为ICM传代的最佳时刻。小鼠囊胚一般培养12～24 h孵化脱带，24～48 h贴壁，72～96 h为传代的最佳时刻。牛胚胎贴附于饲养层上生长，极易从饲养层上剥离；小鼠胚胎镶嵌在饲养层中，滋养层细胞与饲养层细胞间连接紧密。牛ICM色深发黑，集落隆起程度较低；小鼠ICM呈暗黄色，有呈柱状增殖的趋势。牛滋养层呈网状，小鼠滋养层为较致密的单层薄膜。%The growth behavior differences between bovine embryos and murine embryos were studied on PMEF feeder layer using DMEM+15% NBS+0.1 mmol/L β-mercaptoetheanol+0.1 μmol/L Na2 SeO3+10 μg/mL LIF+10 μg/mL IGF-1 as the media.The results showed as follows:In the culture system,bovine blastocyst hatched affer being cultured for 36-60 h,and attached on the feeder layer after being cultured for96-120 h.After cultured for 120-144 h,the growing ICM were passaged at first.Morulae attached on PMEF feeder layer 12-24 h later than blastocyst for passing on blastula stage and hatching period.Murine blastocyst hatched after being cultured for 12-24 h,and attached on the feeder layer after cultured for 24-48 h.After cultured for 72-96 h,the growing ICM were passaged.Bovine embryo lived on the feeder layer and easy to be stripped from the feeder layer,while murine embryo implanted in the feeder layer,and murine trophoblastic cells were closely connected with feeder layer cells arrounding them.The color of bovine ICM was more black than murine ICM.However,the colonies of bovine ICM were lower compared with murine ICM colonies,and murine ICM tended to be column-like.Bovine trophoblast was net-like,and murine trophoblast formed a membranous structure.