桂科Ⅰ号成年公猪体细胞核移植胚胎的体外生产

In vitro Production of Somatic Cell Nuclear Transfered Embryos Derived from Adult Boar of Guike No.1

目前,关于桂科Ⅰ号猪体细胞核移植胚胎体外发育的研究尚未见报道。本研究结果表明,桂科Ⅰ号成年公猪来源的皮肤成纤维细胞(adult boar fibroblast cell,ABFC)和刚出生小公猪来源的皮肤成纤维细胞(newborn boar fibroblast cell,NBFC)对体细胞核移植(somatic cell nuclear transfer,SCNT)胚胎的体外发育没有明显影响;在体外环境培养下,一种组蛋白去乙酰化酶抑制剂—Scriptaid(SCR)对桂科Ⅰ号公猪SCNT胚胎的囊胚率具有促进作用。研究进行3个实验,实验一结果表明,细胞呈现典型的成纤维细胞形态,且生长呈现S型;实验二结果表明,通过将ABFC与NBFC注射到去核的卵母细胞制作SCNT胚胎,ABFC和NBFC来源的桂科Ⅰ号公猪SCNT胚胎的融合率、分裂率、囊胚率和囊胚细胞数没有明显的差异(53.2%vs 61.7%,p>0.05;72.3%vs 75.7%,p>0.05;11.9%vs 11.7%,p>0.05;60.7 vs 57.5,p>0.05);实验三结果表明,通过500 nmol/L SCR处理ABFC来源的SCNT胚胎,与对照组相比,处理组的分裂率和囊胚细胞数没有明显的差异(82.6%vs 80.1%,p>0.05;42.9 vs 41.4,p>0.05);但添加500 nmol/L SCR的ABFC来源的SCNT胚胎的囊胚率明显比对照组的高(21.6%vs 11.5%,p<0.05)。数据表明桂科Ⅰ号成年公猪体细胞可作为供体细胞生产克隆胚胎,并且在体外环境培养下,通过500 nmol/L SCR处理极大地提高克隆胚胎的囊胚率,本研究可为下一步生产存活的桂科Ⅰ号SCNT猪奠定基础。

So far, there were no reports regarding the study about somatic cell nuclear transfer （SCNT） embryos derived from Guike No. 1 in vitro. In this study, there was no significant difference on in vitro development of SCNT embryos derived from adult boar fibroblast cell （ABFC） compared to newborn boar fibroblast cell （NBFC） of Guike No. 1. A histone deacetylase inhibitor-scriptaid （SCR） have a positive impact on blastocyst rate of SCNT in vitro embryos derived from Guike No. 1. Three experiments were performed with routine production of cloned embryos derived from ABFC and NBFC of Guike No. 1. Results showed that： in experiment 1 ： assessment of cell style showed that ABFC and NBFC are very pure fibroblasts, and cell growth curves showed a typical ＂S＂ shape; in experiment 2： we made SCNT embryos via injecting ABFC and NBFC into denucleated oocytes, there was no significant difference in fusion rate, cleavage rate, blastocyst rate and blastocyst cells number （53.2% vs 61.7%, p 〉0.05; 72.3% vs 75.7%, p 〉0.05; 11.9% vs 11.7%, p 〉0.05; 60.7 vs 57.5, p 〉0.05）; in experiment 3： treating embryos derived from ABFC with 500 nmol/L SCR, compared with control group, no significant difference was observed in cleavage rate and blastocyst cells number （82.6% vs 80.1%, p〉0.05; 42.9 vs 41.4, p〉0.05）, but treat- ment with 500 nmol/L SCR in blastocyst rate is more higher than the control group （21.6% vs l 1.5%, p〈0.05）. The data suggested that cells derived from ABFC of Guike No. 1 can make as donor cells for cloned embryos, and extremely improved blastocyst rate of cloned embryos in vitro was observed by treatment with 500 nmol/L SCR. This study laid the foundation for next step that produce alive SCNT pig of Guike No. 1.