一种血清替代物对MDCK细胞培养特性的研究

Study on Characteristics of a Serum Substitute for MDCK Cell Culture

为评估一种血清替代物(Billion cell)支持动物细胞体外培养的效果,分别在含10%Billion cell(3个储存温区37℃、4℃、-20℃)和10%FBS的DMEM培养液中连续传代培养MDCK细胞,利用传代稳定性观察、克隆形成率、绘制生长曲线及生长动力学分析的方法,评价Billion cell对MDCK细胞促生长效果的影响.结果显示:静置培养,-20℃、4℃储存温区的Billion cell和FBS能较好促进MDCK细胞增殖,平均集落形成率从高到低依次为-20℃Billion cell、FBS、4℃Billion cell、37℃Billion cell;生长动力学分析发现平均比生长速率、平均倍增时间和最大增殖浓度从高到低依次为-20℃Billion cell、FBS、4℃Billion cell、37℃Billion cell,故-20℃储存温区的Billion cell支持MDCK细胞培养效果最佳且优于FBS.通过Billion cell对MDCK细胞培养效果影响的研究,旨在开发替代牛血清支持动物细胞培养的有效血清替代物,为生物制品生产提供必要条件.

To assess the effect of a serum substitute(Billion cell)supporting animal cell culture in vitro,MDCK cells were cultured continuously,respectively in containing 10%Billion cell(in storage temperature of 37℃,4℃,20℃)and 10%FBS DMEM medium,and passage stability,cloning efficiency,growth curve and growth kinetics were analyzed.The results showed Billion cell(in storage temperature zones of-20℃and 4℃)and FBS could promote cell growth and proliferation in static culture,and the average colony formation rate was-20℃Billion cell,FBS,4℃Billion cell and 37℃Billion cell from high to low.Growth kinetics analysis showed that the average specific growth rate,average doubling time and maximum proliferation concentration were-20℃Billion cell,FBS,4℃Billion cell and 37℃Billion cell from high to low.By studying the effect of Billion cell on MDCK cell culture,the aim was to explore an effective serum substitutes for bovine serum to support animal cell culture and provide necessary conditions for the production of biological products.