摘要
为建立静置及微载体悬浮培养中国仓鼠卵巢细胞(Chinese hamster ovary cell,CHO)的最适血清的快速筛选方法,在含10%不同批次国产新生牛血清的F12培养液中培养CHO细胞,分别用传代稳定性观察、克隆形成率、绘制微载体培养生长曲线及生长动力学分析的方法,评价不同批次国产新生牛血清对CHO细胞促生长效果的影响.结果显示:静置培养,6个批号新生牛血清均能较好促进CHO细胞增殖,平均集落形成率从高到低依次为D、F、B、E、C、A血清组,且D、F、B组显著高于E组,A组显著低于E组(P<0.05);微载体悬浮培养,6个批号新生牛血清均能实现CHO细胞的微载体悬浮培养,对比发现B、D、F明显优于A、C,E最差.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D、F、B、E、C、A血清组,且D、F组显著高于E组(P<0.05).通过不同批次国产新生牛血清对CHO细胞培养效果影响的研究,旨在为生物制品生产中CHO细胞扩大培养和短时间内成功筛选出培养CHO细胞的最佳血清提供一定的参考依据.
To establish a rapid screening method for the optimal serum for static and microcarrier suspension culture of Chinese hamster ovary cell(CHO),CHO cells were serially sub-cultured in F12 medium containing 10%different groups of domestic newborn bovine serum,and observed by passage stability,cloning efficiency,growth curve and growth kinetics were analyzed.The results showed that all six groups of serum could promote cell growth and proliferation in static culture.In static culture,the orders of the average colony formation rates were D,F,B,E,C and A serum groups from high formation rate to low formation rate.The colony formation rates of D,F and B groups were significantly higher than that of E group(P<0.05).In microcarrier suspension culture,B,D and F groups can obviously promote the growth of proliferation CHO cell,which cell state was obviously better than those of the A and C control groups with the worst in E group.The growth dynamics analysis found that the average growth rate,doubling time and the largest proliferation concentration were D,F,B,E,C,A serum groups from high to low.D,F group was significantly higher than E group(P<0.05).By studying the effects of different types of domestic newborn bovine serum on the culture of CHO cells,the aim was to provide a scientific basis for the expansion of CHO cells in bioproducts production and the successful screening of the best serum for CHO cells in a short period of time.
作者
马春英
王美皓
马花
佛生福
乔自林
马忠仁
王家敏
MA Chun-ying;WANG Mei-hao;MA Hua;FO Sheng-fu;QIAO Zi-lin;MA Zhong-ren;WANG Jia-min(Gansu Tech Innovation Center of Animal Cell,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,China;Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030 China;Life Science and Engineering College,Northwest Minzu University,Lanzhou,730030,China;Lanzhou Bailing Biotechnology Co.LTD,Lanzhou 730010,China)
出处
《西北民族大学学报（自然科学版）》
2021年第2期67-73,共7页
Journal of Northwest Minzu University(Natural Science)
基金
教育部动物医学生物工程创新团队(IRT_17R88)。
作者简介
马春英,女,硕士研究生,主要研究方向为动物细胞培养工程;通讯作者:王家敏,女,讲师,硕士,主要从事动物细胞工程方面的研究.
