用胶原酶消化法培养德保矮马耳缘组织成纤维细胞初探  

A Preliminary Research on Debao Pony Ear Marginal Tissue Fibroblast Cell Cultured by Collagenase Technique

作　　者：马月辉[1] 周向梅[1,2] 关伟军[1] 李晗[1] 

机构地区：[1]中国农业科学院畜牧研究所 [2]中国农业大学动物医学院,北京100094

出　　处：《中国农业科学》2005年第6期1282-1288,共7页Scientia Agricultura Sinica

基　　金：国家自然科学基金(30100132);基础性工作重大专项(2001DEA10006)资助

摘　　要：将德保矮马耳缘组织经胶原酶消化法培养成原代细胞,并成功建立起该组织成纤维细胞系。这种细胞贴壁生长,具有密度接触性抑制性质;该方法获得的原代细胞经传代后接种,细胞群体倍增时间(PDT)为35.9h;细胞染色体众数2n=64的细胞数占91.3%～92.8%;乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)同工酶分析,没有其它细胞系污染;细菌、真菌、病毒、支原体检测阴性。该系符合ATCC要求的细胞系鉴定项目,成为保护矮马这一国家重要畜禽品种的宝贵遗传资源,并为相关遗传学研究提供了有效的试验材料。同时得出胶原酶消化培养法比组织块培养法在获得原代细胞速度快;组织块培养法的细胞群体倍增时间(PDT)为48h。The Debao pony ear marginal tissue fibroblast cell line (NDPEM 2/2) was successfully established using collagenase technique. The characterizations of the cell line were identified as follows: the cells were adherent and of density limitation, and the population doubling time (PDT) of cells made with the collagenase technique was 35.9 h. Chromosome analysis showed that the frequency of cell chromosome number to be 2n = 64 was 91.3%～92.8%. Confirmed by isoenzyme analysis, this cell line had no cross-contamination. Tests for microbial contamination from bacteria, fungi, yeasts or mycoplasma were negative. This newly established cell line meets all the standard quality controls of ATCC. It will provide a precious genetic resource for the reservation of the Debao pony breed, as well as an effective experimental material for the genetic studies on Debao ponies. Collagenase technique was compared with primary explant technique in getting the primary cell at the same time, the former is very fast and the population doubling time (PDT ) of the later is 48 h.

关 键 词：胶原酶消化法 组织 矮马 原代细胞 成纤维细胞系 苹果酸脱氢酶 细胞群体 细胞染色体 乳酸脱氢酶 同工酶分析 遗传学研究 培养法 ATCC 遗传资源 畜禽品种 试验材料 接触性 细胞数 支原体 时间 倍增 接种 病毒 

分 类 号：S821[农业科学—畜牧学]