太行黑山羊成纤维细胞系建立与生物学特性研究

The Establishment of Fibroblast Cell Line and Its Biological Characteristic Research in Taihang Black Goat

以太行黑山羊耳缘组织为材料,采用组织块直接培养法和细胞冷冻技术构建了成纤维细胞系,并进行了细胞活力测定生长动力学观察、微生物污染间检测、染色体标本制备、同功酶分析及生物学研究。结果表明:细胞群体倍增时间(PDT)约为24 h;细胞染色体中二倍体(2n=60)占主体,镜检细胞的百分比为98.2%;乳酸脱氢酶(LDH)、苹果酸(MDH)脱氢酶同工酶分析,没有其他物种污染;细菌、真菌、病毒、支原体检测阴性。该细胞库的各项指标均达到ATCC细胞系鉴定标准。此项研究不仅在细胞水平上保存了这一重要畜禽品种的种质资源,而且亦为其基因组和后基因组及体细胞克隆等研究提供宝贵实验材料。同时,此技术平台的建立必将为其他畜禽遗传资源细胞水平的保存提供技术与理论支撑和保障。

Taking Taihangshan Black Goat ear margin as the study materials, we succeeded in establishing a fibroblast cell bank whose content of samples was 58 by the method of explant culture directly and cell cryoconservation technology. The biological analysis results showed： Population Doubling Time （PDT） of cells was approximately 24 h; Diploid account for 76%～88% in cell＇s chromosome; By Isozyme analysis of Lactic Dehydrogenase （LDH）, Malic Dehydrogenase （MDH）, cross contamination had not taken place; The test results of the bacterium, fungi, virus, mycoplasma were negative. The transfection rate was high, and the extro-gene can be effectively expressed. Every indexes of the cell bank met all the standard quality controls of ATCC. It had not only preserved the germplasm resources of this important Taihangshan Black Goat on the cell level but also provided valuable materials for the research of genome, postgenome and somacloning. Meanwhile, the setting-up of this technological platform will offer technology and theory for storing the cell genetic resources of other animals and poulty.