辽宁绒山羊胎儿成纤维细胞系的建立

Establishment of fetal fibroblast cell line in Liaoning cashmere goat

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。

The aim of the present study was to establish the fetus fibmblast cell line of cashmere goats. The transgenic cashmere goats could be obtained by nucleus transplantation. The tissue explants adherent method and cell freezing technology were used in the process of cell separation and purification. The fibroblast cell line S1 and S2 were obtained by using enzymatic digestion method to purify ceils according to the different tolerance of epithelial ceils and fibmblasts to trypsin. The cell were subjected to sex identification, growth curve and karyotype analysis. The re- suits showed that the donor cell SI was female and S2 was male. The cells grew well and were in good condition with nomal karyotype. There were 30 pairs of chromosomes in the nuclear. The results indicated that the fetus fibmblast in vitro culture system of Liaoning cashmere goat was successfully established. The cells could be used for obtaining high yield transgenic cashmere goats by nucleus transplantation.