Cheng G X, Wang Y G, Zhou X G, Du F, Xiao Y D, Xu S F
Mammalian Embryology and Genetics Engineering Laboratory, Institute of Biotechnology, Jiangsu Agricultural College, Yangzhou, China.
Shi Yan Sheng Wu Xue Bao. 1995 Jun;28(2):147-55.
Goat reconstituted embryos (REs) have been produced by electrofusion-mediated nuclear transplantation method. Single cell derived from normal embryos or REs developed to 8-cell morula stage or the inner cell mass (ICM) of early blastocyst stage was used to fuse with enucleated mature egg (26-28 hrs after injection of LRH). According to the results summarized in table 1 and 2, we decided to adopt the method to embed REs in agarose and then transfer into goat oviduct lumen of host mother for 4-6 days in vivo culture. Normal fertilized eggs seem to develop synchronously, but that of REs are not. Tables 4 and 5 reveal that REs and embryos reconstituted successively can develop normally, no significant difference was found among their development rates. All these experimental results indicate that nuclei of some blastomeres from normal embryos or REs (derived from eight cells to morula or ICM) retained their totipotency for further development. These nuclei can be reprogrammed in host ooplasm, and developed to term.
山羊重构胚(REs)已通过电融合介导的核移植方法产生。取自正常胚胎或重构胚的单细胞发育至8细胞桑椹胚阶段或早期囊胚阶段的内细胞团(ICM),用于与去核成熟卵(注射促黄体素释放激素后26 - 28小时)融合。根据表1和表2总结的结果,我们决定采用将重构胚包埋于琼脂糖中,然后移植到受体母羊输卵管腔中进行4 - 6天体内培养的方法。正常受精卵似乎能同步发育,但重构胚则不然。表4和表5显示,重构胚和相继重构的胚胎能够正常发育,它们的发育率之间未发现显著差异。所有这些实验结果表明,来自正常胚胎或重构胚(源自8细胞至桑椹胚或ICM)的一些卵裂球细胞核保留了进一步发育的全能性。这些细胞核可在受体卵质中重编程,并发育至足月。