岷县黑裘皮羊肾组织成纤维细胞系的建立及其生物学特性研究

采集岷县黑裘皮羊肾组织,用胰蛋白酶热消化法制备原代细胞,通过差速消化和差速贴壁法进行继代培养和细胞纯化,并对复苏细胞的形态、活力、生长曲线、荧光蛋白质粒转染表达、核型以及乳酸脱氢酶同工酶等生物学特性进行了分析。结果显示:原代和传代细胞生长形态均好,群体倍增时间为28.3 h;染色体2 n=54,二倍体占主体(84%)乳酸脱氢酶同工酶电泳图谱有明显特征,未见LDH4、LDH5谱带,与其它羊有明显区别;外源性荧光蛋白转染质粒在该细胞中能进行复制和表达;细菌、真菌、病毒、支原体检测呈阴性。表明该研究已成功建立岷县黑裘皮羊肾组织成纤维细胞系,为在细胞水平上保存岷县黑裘皮羊种质资源以及进行相关研究提供了理想的生物材料。

Establishment and Characteristics of Minxian Black Fur Sheep Kidney Explant Fibroblast

The Minxian black fur sheep kidney explant was cultured with trypsinization and then subcultured.Observations and analysis on the morphology,dynamic growth,fluorescin plasmid transfection and expression,karyotype,isoenzyme of lactate dehydrogenase were carried out.The results showed that the population doubling time of cells(PDT) was 28.3 h,the chromosomes were 54 and diploid cells were dominant of 84%.The banding patterns of the isoenzyme of lactate dehydrogenase had significant difference.As follow,the exogenous plasmid could copy and express in the cell,tests for bacteria,fungi and yeasts,virus and mycoplasma were negative.These suggested that the cell line was successfully estabiished,and it made the Minxian black fur sheep,a national important genetic resource,preserved in cell level,as well as an ideal experimental material for genetic studies.