| 贵德黑裘皮羊耳成纤维细胞系的建立和生物学特性的研究 |
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| 引用本文： | 陆会宁,刘丑生,王志刚,张利平,赵俊金,孟飞.贵德黑裘皮羊耳成纤维细胞系的建立和生物学特性的研究[J].中国草食动物,2007,27(2):3-6. |
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| 作者姓名： | 陆会宁 刘丑生 王志刚 张利平 赵俊金 孟飞 |
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| 作者单位： | 1. 甘肃农业大学动物科技学院,兰州,730070
2. 全国畜牧兽医总站畜禽牧草种质资源保存利用中心,北京,100094 |
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| 基金项目： | 农业部畜禽牧草品种资源保护项目;科技部农业科技成果转化基金 |
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| 摘 要： | 以贵德黑裘皮羊耳缘组织为材料，采用组织块贴壁培养法和细胞冷冻技术成功构建了成纤维细胞系，并对培养细胞进行了形态学、生长动力学观察、细胞活力测定、核型分析和微生物检测。结果表明：细胞群体倍增时间（PDT）约为36h，冻存前细胞活力为96．2％，以DMEM／F-12＋20％FBS中添加10％DMSO冻存成纤维细胞，解冻后细胞活力为93．6％，24h后的贴壁率为85％。在传代10次之前，细胞染色体中二倍体（2n=54）占主体，约为82％～90％，细菌、真菌、病毒、支原体检测为阴性。该细胞系的建立，使贵德黑裘皮羊这一国家重要种质资源在细胞水平上保存下来，也为体细胞克隆等研究提供了理想的生物材料。
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| 关 键 词： | 贵德黑裘皮羊 成纤维细胞系 细胞培养 生物学特性 |
| 文章编号： | 1007-9726(2007)02-0003-04 |
| 修稿时间： | 2006年10月30 |
The Establishment of Fibroblast Cell Line and Its Biological Characteristic Research in Guide Black fur sheep |
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| Lu Hui-ning,Liu Chou-sheng,Wang Zhi-gang.The Establishment of Fibroblast Cell Line and Its Biological Characteristic Research in Guide Black fur sheep[J].China Herbivores,2007,27(2):3-6. |
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| Authors: | Lu Hui-ning Liu Chou-sheng Wang Zhi-gang |
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| Institution: | Lu Hui-ning1,Liu Chou-sheng2,Wang Zhi-gang2 |
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| Abstract: | Taking Guide Black fur sheep ear margin as the study materials.we succeeded in establishing a fibroblast cell line by the method of explant adherence culture and cell cryoconservation technology.Observations on morphology,dynamic growth,determination of viability,analysis of karyotype,test of microorganism were carried out.The results showed :Population Doubling Time(PDT)of cells was approximately 36 h,Cell viability before freezing was 96.2%,The fibroblasts were frozen in DMEM/F-12 20?S by adding 10% DMSO,After thawing Cell viability was 93.6%,The attachment rates of thawed cells after 24h was 85%.When cultured up to passage ten,Diploid(2 n=54)account for 82%~90% in cells chromosome.tests for microbial contamination of bacteria,fungi and yeasts,virus and mycoplas ma were negative.The test result of mycoplasma was negative.The newly established cell line not only preserved the gemlplasm resources of this important Guide Black fur sheep on the cell level,but also provided valuable materials for the research of somatic cloning. |
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| Keywords: | Guide Black fur sheep fibroblast cell line cell culture biological characteristics |
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