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.%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 subcul-tured 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 plas-mid (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.
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