牛-兔种间重组胚体外发育能力的研究

Study on Development of Cloned Embryo Using Bovine Somatic Cell and Rabbit Oocyte in vitro

以超排的青年母兔和经产母兔卵母细胞为受体细胞,以处于休眠期和非休眠期的黑毛和牛耳成纤维细胞为核供体,共获得180枚种间重组胚,采用M199+10%FBS和RD+10%FBS2种培养液进行体外培养,有112枚发育到2-细胞期(62.2%),26枚发育到桑椹胚期(14.4%),20枚发育到囊胚期(11.1%)。结果显示,供体细胞处于休眠期与否,对重组胚的卵裂率无显著影响(P<0.05),但以休眠期细胞为核供体的重组胚8-细胞～16-细胞胚的发育率及桑椹胚发育率显著高于非休眠期细胞的重组胚(P<0.05),二者在囊胚发育率上差异极显著(P<0.01);以M199为培养液的胚胎在卵裂率、桑椹胚发育率和囊胚发育率上与RD组无显著差异(P>0.05)。

Total 180 reconstituted embryos were produced by using Japan Black Cattle's ear fibroblast cell whichstaying at G0 or non-G0 cell cycle stage as donor and oocyte superovulated from multiparous or young rabbit as recipient.After cultured in two kinds of medium: M199+10%FBS or RD+10%FBS, 112 of them developed to 2-cell stage (62.2%), 26developed to morula stage (14.4%), 20 developed to blastocyst stage (11.1%). The result indicated that no significantdifference was obsearved in cleavage rate between two groups of reconstituted embryo which using G0-stage cell and non-G0 stage cell as donor respectively, but when used G0-stage cell as donor, 8-cell-16-cell rate and morula rate were raisedsignificantly (P<0.05), blastocyst rate was raised significantly (P<0.01). No significant difference in cleavage rate, morularate and blastocyst rate was obsearved when using two kinds of culture medium (P>0.05).