【摘要】 采用组织块培养法,分别以DMEM(高糖)和DMEM/F12两种合成培养基为基础液,添加15mL·L-1胎牛血清(FBS)为细胞生长液对牦牛耳组织进行原代培养。同时,比较不同冷冻程序对细胞冷冻的影响,探讨牦牛体细胞在体外环境下的生长参数及染色体变化。结果表明,组织块在第6天开始有细胞游离,14天长至单层,两种基础培养基对细胞生长的影响差异不显著(P﹥0.05)。细胞在-20℃短时间(不超过1h)冷冻效果较好,但平衡时间过长对细胞解冻后生长影响差异极显著(P﹤0.01)。染色体分析显示,牦牛成纤维细胞在体外条件下生长,不同传代次数(3代和12代)细胞染色体核型稳定,在所得的中期分裂相细胞中,二倍体细胞都在96/以上。可用于体细胞核移植操作和其他研究。更多还原

【Abstract】 Yak ear fibroblasts were cultured in 150mL·L-1 fetal bovine serum(FBS) with different culture media. The effect of different refrigeration procedure and the change of chromosome of fibroblasts culture in vitro was analys. The result shoused that the cells erupted in 6 days, and confluented together in 14 days. No obvious different effects were found on the differents base media.(P>0.05). Under -20℃ condition, refrigeration time had a siguificant effect(P<0.01) on cell growth. Chromosome karyotype analyze indicated that after 3 and 12 passage in vitro culture no abnormal karytype was found. It could be used for nuclear transfer operation and othe study.更多还原