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为了成功建立绒山羊胎儿成纤维细胞系,利用核移植法获取转基因绒山羊,试验采用组织块贴壁法和细胞冷冻技术,并根据上皮细胞和成纤维细胞对胰蛋白酶的耐受性不同,采用酶消化法纯化细胞获得成纤维细胞系S1和S2,并对该细胞进行性别鉴定、生长曲线测定与核型分析。结果表明:该供体细胞S1为雌性,S2为雄性,其生长旺盛,状态良好,核型正常,有30对染色体。说明试验成功建立了辽宁绒山羊胎儿成纤维细胞的体外培养体系,可用于通过核移植法获取高产绒量的转基因绒山羊。
In order to establish the cashmere goat fetal fibroblast cell line successfully, the transgenic cashmere goat was obtained by the nuclear transfer method. The tissue block adherent method and the cell freezing technique were used in the experiment. According to the different tolerance of the epithelial cells and fibroblasts to trypsin, The cells were purified by enzymatic digestion to obtain fibroblast cell lines S1 and S2, and sex determination, growth curve determination and karyotype analysis of the cells were performed. The results showed that the donor S1 was female and S2 was male. Its growth was good, with good karyotype and 30 pairs of chromosomes. The experiment successfully established in vitro culture system of Liaoning Cashmere goat fetal fibroblasts can be used to obtain high cashmere yield transgenic cashmere goats by nuclear transfer method.