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Establishment and characteristics of a Bactrian camel fetal ear marginal fibroblast cell line

DIAO Ruixin, ZHANG Yong, HU Junjie, CHANG Weihua, LIN Tao, ZHANG Zhiqiang,XU   

  • Online:2012-11-22 Published:2012-11-20

双峰驼胎儿耳缘组织成纤维细胞系的建立及其生物学特性鉴定

刁睿鑫 张勇 胡俊杰 常卫华 林涛 张志强   

  1. 甘肃农业大学动物医学院

Abstract: The experiment was conducted to establish a Bactrian camel (Camelus bactrianus)fetal ear marginal fibroblast cell line. The Bactrian camel fetal ear marginal tissue was cultured with explants or trypsinization,which then was subcultured and purified through trypsin digestion and differential anchoring velocity. Analysis on cell morphology,cell viability,
growth dynamics,immunocytochemistry,karyotype,transfection and expression of green fluorescent protein marker plasmid (GFP),and microbial contamination were carried out. The results showed that the population doubling time (PDT) of the cells was 47 2 h;the majority chromosome number was 74;the immunocytochemistry was positive for vimentin and negative for cytokeratin;an exogenous plasmid could integrate and express in the cells;detection for microbial contamination was negative. As a consequence,we succeeded in establishing the fibroblast cell line of Bactrian camel on cell level which is a significant national germplasm resource,providing ideal experimental material for future studies of the gene bank and somatic cell cloning,etc.

Key words: Bactrian camel (Camelus bactrianus), Biological characteristics, Ear marginal tissue, Fibroblast cell line

摘要: 本研究旨在建立双峰驼胎儿耳缘组织成纤维细胞系。采集双峰驼胎儿耳缘组织,用植块培养法和消化分离法对其进行原代培养,差速消化和差速贴壁法进行继代培养和细胞纯化,并对培养细胞的形态、细胞活力、生长动力学、免疫组织化学、核型、绿色荧光蛋白基因质粒(GFP)转染表达以及微生物污染等生物学特性进行了分析。结果表明,原代和传代细胞形态正常,群体倍增时间(PDT)为47. 2 h,细胞生长较快;不同代数的染色体众数为2n = 74;免疫组织化学鉴定波形蛋白抗体呈阳性,角蛋白抗体呈阴性;外源质粒能在该细胞内整合和正确表达;微生物污染检测呈阴性。因此该研究已成功建立了双峰驼胎儿耳缘组织成纤维细胞系,在细胞水平上保存了双峰驼这一国家重要种质资源,为基因组文库和体细胞克隆等研究提供了理想的生物材料。

关键词: 双峰驼, 耳缘组织, 成纤维细胞系, 生物学特性