信阳水牛成纤维细胞的体外培养及生物学特性分析

Establishment of Fibroblast Line and Its Biological Characteristics in Xinyang Buffalos

为建立信阳水牛成纤维细胞系,以信阳水牛耳缘组织为实验材料,采用组织块培养法进行原代和传代培养,并对其形态学、生长曲线、细胞活力、染色体、微生物检测等生物学特性进行分析。结果表明,培养的细胞形态为典型的成纤维细胞,生长曲线呈"S"型,细胞冻存前和复苏后的活率均在95%以上,细胞在10代之内二倍体均在96%以上;细菌、真菌、支原体检测结果为阴性。信阳水牛成纤维细胞的体外培养可在细胞水平上对信阳水牛的遗传资源进行保存,也可为体细胞克隆提供实验材料。

To establish the fibroblast line of Xinyang buffalos,Xinyang buffalo ear margin was used as the study material,the primary fibroblasts and subculture were made by the method of explant culture directly.Observations on morphology,dynamic growth curves,viability,karyotype,microorganisms and other characteristics were detected.The results showed that the cell morphology was typical fibroblasts,the cell growth curve was a type of“S”,cell viability was above98%before freezing and after thawing.Diploid cells was more than96%before10generations.Tests of the bacteria,fungi and mycoplasma were negative.The newly established cell line made a national important genetic resource preserved in cell level,as well as provided an ideal experimental material for the somatic cell cloning of Xinyang buffalos.