To develop a method of rabbit ESCs-like ( embryonic stem cells like) culture in vitro for obtaining the rabbit ESCs (embryonic stem cells) line in future, not only the rates of inner cell mass (ICM) proliferation, formation and positive alkaline phosphatase (AKP) of the primary rabbit ICM colonies from in vitro and in vivo were comparativly studied on different feeders, but also rabbit ESCs-like were passage cultured separately on MEF feeder and REF feeder. The results showed that there were significantly higher rates of development and attachment of blastocyst from morula without zona pellucida than that with zona pellucida ( P ＜ 0. 05 ). The positive AKP rate of primary ICM colonies in ectogenesis blastula was significantly lower than that in vivo( P ＜0. 05). Compared to MEF feeder group, the rate of primary colonies formation was lower remarkably on REF feeder (P ＞0.05), but the rate of positive AKP of the primary colonies was higher significantly than others ( P ＜0.05). Although ESCs-like could be passage cultured on both of REF and MEF feeders, the rate of ESCs-like colonies formation following F3 in MEF feeder group was higher significantly than that in the REF feeder group ( P ＜ 0. 05 ).%为了确立兔类胚胎干细胞(ESCs-like)体外培养方法,研究探讨了兔囊胚形成环境与饲养层细胞对内细胞团(ICM)增殖、ICM原代集落形成以及集落碱性磷酸酶(AKP)阳性率的影响,并在MEF和REF饲养层上进行了兔ESCs-like的体外培养.结果显示,去除透明带桑葚胚在体外具有较高的囊胚发育率和囊胚贴壁率(P＜0.05),体外发育囊胚ICM原代集落的AKP阳性率却显著低于体内发育囊胚(P＜0.05);与MEF饲养层相比,REF饲养层上ICM原代集落的形成率显著偏低(P＞0.05),但原代集落的AKP阳性率却显著高于其余各组(P＜0.05);MEF饲养层与REF饲养层均支持兔ESCs-1ike的传代培养,但F3代后MEF组中ESCs-like的集落形成率显著高于REF组(P＜0.05).
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