Modliński J A
J Embryol Exp Morphol. 1980 Dec;60:153-61.
Haploid embryos were obtained by microsurgical removal of one pronucleus, followed by doubling of the haploid chromosome set with Cytochalasin B (CB), either at the first or second mitosis. This procedure provides a source of fully homozygous diploid embryos which were grown in vitro or in vivo. The effect of CB treatment before and during operation on the course of enucleation and further development of embryos was studied. Out of 81 eggs made diploid at 2-cell stage and transplanted into the oviducts of immature or pseudopregnant recipients 27 morulae and blastocysts were recovered, but not a single case of implantation occurred by the eighth or ninth day of development. After 72-80 h of in vitro culture, most of the homozygous embryos were morulae but after an additional 24 h the majority of them transformed into blastocysts. The rate of development of homozygotes was markedly better than that of haploids, which progressed beyond morula stage. The immediate survival rate of operated eggs was dependent on whether or not the eggs were pre-incubated and the enucleation was performed in the presence of CB. In the former case the immediate survival rate was nearly twice as high as in the absence of CB, but more of the treated eggs underwent fragmentation and early developmental arrest.
通过显微手术去除一个原核获得单倍体胚胎，然后在第一次或第二次有丝分裂时用细胞松弛素B（CB）使单倍体染色体组加倍。该程序提供了完全纯合的二倍体胚胎来源，这些胚胎在体外或体内生长。研究了手术前和手术过程中CB处理对去核过程和胚胎进一步发育的影响。在81个在2细胞阶段变为二倍体并移植到未成熟或假孕受体输卵管中的卵中，回收了27个桑葚胚和囊胚，但在发育的第八天或第九天没有发生一例着床。体外培养72 - 80小时后，大多数纯合胚胎为桑葚胚，但再过24小时，它们中的大多数转化为囊胚。纯合子的发育率明显优于单倍体，单倍体超过桑葚胚阶段的进展情况较好。手术处理的卵的即时存活率取决于卵是否预先孵育以及去核是否在CB存在的情况下进行。在前一种情况下，即时存活率几乎是不存在CB时的两倍，但更多经处理的卵发生了碎片化和早期发育停滞。
