摘要
兰州大尾羊是中国濒临灭绝的物种,通过体细胞培养和长期保存,在细胞水平上保存珍贵的种质资源。采用胰蛋白酶消化分离并培养原代肾细胞和睾丸细胞,并通过差速消化和差速贴壁法纯化成纤维细胞并用液氮保存,细胞复苏后进行活力、形态、生长特性、微生物污染和染色体等检查,保存兰州大尾羊26个个体的肾细胞和9个个体睾丸细胞,保存细胞生长良好,均呈成纤维型,平均复苏活力为98.6%和95.2%,生长曲线均呈"S"型,最大增殖密度分别为3.15×105 mL-1和2.92×105 mL-1,倍增时间分别为29.3h和30.8h,细菌、真菌、病毒和支原体检查均为阴性,染色体为2n=54,二倍体占88%和90%,说明成功分离培养并保存兰州大尾羊肾和睾丸组织细胞。
In China,Lanzhou fat-tailed sheep is endangered species. This rare resource was preserved in cell level by culturing and preserving the cells of this species. The kidney and testicles explants of Lanzhou Fat-tailed Sheep were cultured with trypsinization method. The primary cells were purified and preserved in liquid nitrogen. Observations on vitality, morphology, growth characteristic, microbi- al contamination and analysis of karyotype were carried out. The kidney cells from 26 individuals and testicle cells from 9 individuals were conserved. They were fibroblasts and grew well; the vitalities of these two kinds of recovery cells were 98. 6% and 95.2%; the growth curve showed ＂S＂ type; the maximum proliferation densities were separately 3.15×10^5 mL 1 and 2.92 ×10^5 mL^-1 ; the population doubling time of cells （PDT） were 29.3 h and 30.8 h,respectively; the test results for bacteria,fungi, virus and mycoplasma were all negative; the chromosomes were 54 and diploid cells were dominant. These showed that the kidney and testicle cells from Lanzhou fat-tailed sheep were isolated and cul- tured successfully, which makes the important genetic resource preserved in cell level.
出处
《西北农业学报》
CAS
CSCD
北大核心
2013年第4期17-21,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
国家科技基础平台项目(2005DKA21101-29)
兰州市科技计划项目(2006-2-59)
关键词
兰州大尾羊
肾细胞
睾丸细胞
培养
保存
Lanzhou fat-tailed sheep
Kidney cell
Testicle cell
Cell culture
Cell preservation
