兰州大尾羊耳缘组织成纤维细胞系的建立

Establishment of Lanzhou Fat-tailed Sheep Marginal Ear Fibroblast

通过物种体细胞培养和长期保存,可在细胞水平上保存珍贵的种质资源。采用组织块培养法培养耳缘组织原代细胞,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,细胞复苏后进行了活力、形态、生长特性、微生物污染和染色体等检查。结果表明:保存了兰州大尾羊27个个体的耳缘细胞,保存的细胞平均复苏活力97.6%,生长良好,形态呈成纤维型,生长曲线呈"S"型,最大增殖浓度3.01×105个.mL-1,倍增时间为26.1h,细菌、真菌、病毒和支原体检查为阴性,染色体为2n=54,二倍体占88%。最终,成功培养并保存了兰州大尾羊耳缘组织成纤维细胞,使这一珍贵的种质资源在细胞水平上得以长期保存。

It can preserve rare resources at cell level through culturing and preserving cells of the species.The marginal ear primary cells were cultured by tissue pieces method.And the primary cells were purified and preserved with liquid nitrogen.Observations on dynamic growth,morphology,growth characteristic,microbial contamination and analysis of karyotype were carried out.The results showed that the cells from 27 sheep were preserved.The ear cells were fibroblast and grew well;the vitality was 97.6%;the growth curve showed ＇S＇ type;the maximum proliferation concentration was 3.01×105 cells·mL-1;the population doubling time（PDT）was 26.1 h;the tests for bacteria,fungi,virus and mycoplasma were all negative;and the chromosomes were 54 and diploid cells were dominant for 88%.These showed that the cell line was established successfully,which making the important genetic resource preserved in cell level for long term.