不同传代比例对MDCK细胞培养效果的影响  

Effect of subculture ratio on culture of MDCK cells

作　　者：王美皓 李自良 赵彩红 李倬[2] 张雪梅[3] 乔自林[1] 马忠仁[1] 王家敏[1] WANG Mei-hao;LI Zi-liang;ZHAO Cai-hong;LI Zhuo;ZHANG Xue-mei;QIAO Zi-lin;MA Zhong-ren;WANG Jia-min(Gansu Tech Innovation Center of Animal Cell,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,Gansu Province,China)

机构地区：[1]西北民族大学生物医学研究中心,甘肃省动物细胞技术创新中心,甘肃兰州730030 [2]西北民族大学生命科学与工程学院,甘肃兰州730030 [3]长春生物制品研究所,吉林长春130012

出　　处：《中国生物制品学杂志》2020年第11期1230-1234,1239,共6页Chinese Journal of Biologicals

基　　金：国家科技重大专项重大新药创制项目(2015ZX09102016);西北民族大学动物细胞培养与代谢工程创新团队(319201-90004);甘肃省重点研发计划(18YF1FA121)。

摘　　要：目的探讨不同传代比例对MDCK细胞培养效果的影响。方法用含10%牛血清的DMEM培养液培养MDCK细胞,按不同传代比例(1∶4、1∶5、1∶6、1∶7、1∶8)连续传5代,观察细胞形态、长势、致密程度,绘制生长曲线,并通过生长动力学方法评价不同传代比例对MDCK细胞培养效果的影响。同时按最佳传代比例于细胞工厂(1层、10层)中进行扩大培养。结果不同传代比例的MDCK细胞均可在48 h增殖至致密单层,细胞生长曲线均趋于"S"型,细胞最大增殖密度均达11×10^4个/cm^2以上。传代比例为1∶8的MDCK细胞平均最大比生长速率显著高于1∶4、1∶5、1∶6的比例(P<0.05),倍增时间显著短于1∶4、1∶5、1∶6、1∶7的比例(P<0.05)。按1∶8的比例在1层和10层细胞工厂中连续传代培养的MDCK细胞生长良好,且稳定。结论按1∶8比例传代的MDCK细胞培养效果最佳。Objective To investigate the effect of different subculture ratios on the culture of MDCK cells.Methods MDCK cells were cultured in DMEM medium containing 10%bovine serum and subcultured at different subculture ratios(1∶4,1∶5,1∶6,1∶7 and 1∶8)for five passages.The morphology,growth and density of cells were observed,and the growth curve was plotted,based on which the effect of subculture ratio on culture efficiency of MDCK cells was evaluated by growth kinetics analysis.The cells were subjected to expanded culture in cell factories(one and ten layers)at the optimal subculture ratio.Results MDCK cells at different subculture ratios were proliferated to dense monolayer 48 h after subculture,of which the growth curve tended to a S shape,and the maximum proliferation density reached more than 11×10^4 cells/cm^2.The average maximum specific growth rate of MDCK cells at a subculture ratio of 1∶8 was significantly higher than those at 1∶4,1∶5 and 1∶6(P<0.05),while the population doubling time was significantly shorter than those at 1∶4,1∶5,1∶6 and 1∶7(P<0.05).The MDCK cells cultured in one and ten layer cell factories at a subculture ratio of 1∶8 grew well and stably.Conclusion The subculture ratio of 1∶8 was suitable for culture of MDCK cells.

关 键 词：MDCK细胞 传代培养 生长曲线 

分 类 号：R373.23[医药卫生—病原生物学] Q75[医药卫生—基础医学]