藏羚与几种牛科动物成纤维细胞的特性比较

Comparative of the cytogenetic characteristics of Pantholops hodgsonii and other three species in Bovidae

为研究高原低氧环境对哺乳动物细胞生长特性的影响,本文以高原低氧动物藏羚和同科的阿尔巴斯白绒山羊、蒙古绵羊及鲁西黄牛的耳源组织细胞为材料,利用DMEM/F12培养液进行培养,建立了4种动物的体外培养成纤维细胞系,并对其进行形态学观察、生长曲线绘制、凋亡检测、核型分析等生物学特性检测。结果表明:4种动物的细胞经差异贴壁法联合消化排除法,可获得纯化的成纤维细胞,细胞形态一致且活力稳定,4种动物细胞的形态与生长曲线无明显差异;藏羚、绒山羊、绵羊、牛同代次细胞生长倍增时间分别为26 h、22 h、23 h和22 h;经流式细胞仪检测,细胞凋亡率分别为5.5%、2.7%、8.6%和1.5%;藏羚、绒山羊、绵羊与牛的染色体数目分别为60、60、54和60。藏羚与绒山羊、牛染色体数目相同,且除性染色体外,其余均为端部着丝粒类型,而绵羊存在6条中着丝粒类型染色体。该结果表明,在体外培养条件下,藏羚的体细胞生长特性与同科的其他物种的细胞生长特性存在明显差异。这为从细胞水平上研究藏羚对环境低氧适应的分子机理提供了基础。

The ear tissues of four bovidae species Pantholops hodgsonii, Capra hircus, Ovis aries and Bos taurus were collect- ed and cultured in vitro with DMEM/F12 media and the cell lines were established. The cell morphologies, growth curves, apoptosis and karyotypes were investigated. The results showed that fibroblast cells from the four different species could be cultured and purified by the combination of differential attachment protocol and digestion elimination methods. The cells had consistent morphology and kept stable vitality. The cell morphologies and growth curves were with not different among the four different species＇ cells. The population doubling times （PDT） of Pantholops hodgsonii, Capra hircus , Ovis aries and Bos tau- rus were 26 h, 22 h, 23 h and 22 h, respectively. The cell apoptosis rates were 5.5%, 2. 7%, 8.6% and 1.5%, re- spectively, when assayed by the flow cytometry. The chromosome number of Pantholops hodgsonii, Capra hircus, Ovis aries and Bos taurus are 60, 60, 54 and 60 respectively. Karyotype analyses showed that Ovis aries have 6 metacentric chromo- somes, and those of all of the other species are telocentric type except the bovine X chromosomes are metacentric. These re- suits indicated that the cell growth characteristics of Pantholops hodgsonii had obvious differences when compared to the other Bovidae species. Our research provides a basis to study the molecular mechanisms of Pantholops hodgsonii adaptation to hy- poxia.