山羊转t-PA突变体基因的体细胞核移植研究

The Study of Matic Cell Nucear Tranfer of Transgenin (t-PA) in Goat

比较了用转t-PA突变体基因的胎儿成纤维细胞和卵丘细胞作供体时核移胚的发育情况。结果表明:卵丘细胞作供体时,核移胚的卵裂率(82.9%)和桑葚胚率(55.9%)均高于胎儿成纤维细胞卵裂率(75.7%)和桑葚胚率(48.7%),但差异不显著(P>0.05);卵丘细胞作供体时的囊胚率(19.7%)则高于胎儿成纤维细胞(11.0%),差异显著。(P<0.05)。比较了饥饿处理与否对胎儿成纤维细胞作供体时转基因核移胚的发育情况。结果表明:供体细胞饥饿后,其核移胚的卵裂率(75.6%)与供体不饥饿的卵裂率(75.8%)差异不显著;供体细胞饥饿后,其核移胚的桑葚胚率(46.5%)和囊胚率(11.0%)高于供体不饥饿的桑葚胚率(39.3%)和囊胚率(9.0%),但差异不显著。比较了SOFaa和CR1aa分别添加10%FBS和bFF后,转基因核移胚的发育情况。结果表明:用CR1aa液培养时,添加10%bFF,其囊胚率(9.1%)高于添加10%FBS(4.2%)(P>0.05);用SOF液培养时,添加10%bFF,其囊胚率(11.2%)高于添加10%FBS(5.5%)(P>0.05)。说明用SOF液优于CR1aa液。

The development of nuclear transfer embryos derived from fetal fibroblasts including t-PA gene and ovary granulose cells were compared. The cleavagetate (82.9%)and Morulae rate (55.9%)of ovary granulose cells were more high than those of fetal fibroblasts(75.7%,48.7%). Blastocyst rate (19.7%) of ovary granulose cells was significantly higher than that of fetal fibroblasts (11.0%). The development of nuclear transfer embryos derived from fetal fibroblasts including t-PA gene in media DMEM/F12 including 0.5%FBS for starvation and including 10%FBS were compared. The cleavagetate(75.6%) of nuclear transfer embryos in media DMEM/F12 adding 0.5%FBS is not significantly defferenter than that (75.8%)in media DMEM/F12 adding10%FBS. Morular rate (46.5%) and blastocyst rate (11.0%) in media DMEM/F12 including 0.5%FBS were higher than those (39.3%, 9.0%) in media DMEM/F12 adding10%FBS (P>0.05). The blastocyst rate(9.1%) nuclear transfer embryos in media SOF adding 10% bFF was higher that(4.2% )10% FBS. The blastocyst rate(11.2%) nuclear transfer embryos in media SOF adding 10% bFF was higher that(5.5% )10% FBS. SOF was better than CR1 for the early development in nuclear transfer embryos.