Vero细胞的生长代谢特征及动力学模型构建  

Metabolic characteristics and kinetic model of Vero cell

作　　者：平玲 顾朝剑 吴易梅 张立敏 陈溢娟 杨净思 沈武玲 PING Ling;GU Chaojian;WU Yimei;ZHANG Limin;CHEN Yijuan;YANG Jingsi;SHEN Wuling(Institute of Medical Biology,Chinese Academy of Medical Sciences,Kunming 650032,China)

机构地区：[1]中国医学科学院昆明医学生物学研究所,云南昆明650032

出　　处：《生物技术》2024年第4期448-453,共6页Biotechnology

基　　金：云南省创新疫苗技术与产业转化研发平台(202002AA10009,202302AA310003,202401AT070147)。

摘　　要：[目的]解决Vero细胞在大规模高密度培养过程中营养物质耗尽,代谢产物积累导致细胞生长受限的问题。[方法]固定床生物反应器培养Vero细胞,通过换液培养和灌流培养两种方式比较细胞增殖、营养物质的消耗、代谢产物的生成,基于Logistic方程、Luedeking-Piret方程建立动力学模型,并根据实际检测数据,应用Origin软件进行非线性拟合。[结果]换液培养和灌流培养两种培养方式培养Vero细胞,两种培养方式达到的最大细胞密度分别是4.35×10^(6)/mL和4.34×10^(6)/mL,在对数生长期,细胞的生长速率分别是0.54 d^(-1)和0.67 d^(-1),葡萄糖的最大消耗量分别是14.95 g/d和17.1 g/d,谷氨酰胺的最大消耗量分别是2.725 g/d和3.7 g/d;Origin软件进行非线性拟合,得到的建模参数通过验证两种培养方式的细胞生长模型预测值与实验值拟合度较高。[结论]大规模培养Vero细胞,利用灌流培养的细胞生长速率较快,对营养物质的利用率较高,是理想的细胞培养方式;所建立的代谢动力学模型能较好的反应细胞在生长过程中的反应动力学过程。[Objective]To solve the problem of cell growth restriction caused by nutrient depletion and accumulation of metabolites in the process of large-scale culture.[Method]Vero cells were cultured in the fixed bed reactor by liquid replacement and perfusion to compare the proliferation of cells,the consumption of nutrients and the production of metabolites,a dynamic model was established based on the Logistic equation and Luedeking-Piret equation and according to the date gained nonlinear fitting by software Origin.[Result]Vero cells were cultured by liquid replacement and perfusion,and the maximum cell density achieved by the two culture methods were 4.35×10^(6)/mL and 4.34×10^(6)/mL,in the logarithmic growth phase,the cell growth rate were 0.54 d^(-1)and 0.67 d^(-1),the maximum consumption of glucose were 14.95 g/d and 17.1 g/d,and the maximum consumption of glutamine were 2.725 g/d and 3.7 g/d,respectively.The obtained modeling parameters were verified to have a high degree of fitting between the predicted values and the experimental values by software Origin.[Conclusion]In large-scale culture of Vero cells,perfusion culture of Vero cells has a faster growth rate and higher utilization rate of nutrients,which is an ideal cell culture method.The established metabolic kinetics model can reflect the reaction kinetics of cells growth well.

关 键 词：VERO细胞 换液培养 灌流培养 代谢物消耗 代谢物生成 ORIGIN软件 非线性拟合 动力学模型 

分 类 号：Q784[生物学—分子生物学]