CHO工程细胞无血清悬浮分批培养的生长代谢特征及动力学模型

Metabolic characteristics and kinetic model of recombinant CHO cells in serum-free suspension batch culture

摘要:

以悬浮适应的表达尿激酶原CHO工程细胞为研究对象，在100 mL的摇瓶中进行无血清悬浮培养，以细胞密度、细胞活力、Pro-UK活性、葡萄糖比消耗速率 (qglc)、乳酸比生产速率 (qlac)、乳酸对葡萄糖的得率系数 (Ylac/glc) 为观察指标，同时以细胞有血清悬浮培养作为参照，考察CHO工程细胞无血清悬浮培养生长和代谢特征。观察结果表明，CHO工程细胞在无血清及有血清悬浮培养条件下表现为大致相似的细胞生长和代谢特征。在此基础上，依据实际检测的数据，应用MATLAB软件对细胞对数生长期的细胞生长、乳酸生成及葡萄糖消耗的模型参数进行非线性规划，获得全局性收敛的最优参数估计值，建立了细胞在无血清培养条件下的生长及代谢动力学模型。

Abstract:

By using the cell density, cell viability, Pro-UK activity, specific consumption rate of glucose (qglc), specific production rate of lactate (qlac), yield of lactate to glucose (Ylac/glc) and as the evaluation indexes, the growth and metabolism characteristics of pro-urokinase (Pro-UK) expressing CHO cells in serum-free suspension batch culture were examined and compared to those in serum-containing suspension batch culture. We observed hardly differences in growth and metabolism characteristics between the CHO cell populations grown in serum-free suspension batch culture and serum-containing suspension batch culture. The optimal mathematical model parameters for the CHO cells grown in suspension batch culture were obtained by non-linear programming of data representing the growth, substrate consumption and product formation of the CHO cells during logarithmic growth phase using MATLAB software, and the kinetic model of the cell growth and metabolism in serum-free culture were established.