Marc-145细胞无血清培养液组分的优化筛选

目的：对Marc-145细胞无血清培养液组分及浓度进行优化筛选。方法：采用Plackett-Burman设计和中心组合旋转设计对不同的营养成分进行优化筛选。结果：经Plackett-Burman设计，利用20组实验对14种不同营养成分进行考察，发现孕酮、抗氧化试剂、乙醇胺、维生素B12、维生素B12和脂肪酸复合剂对Marc-145细胞无血清悬浮生长的比生长速率有显著影响；针对该6种因素进行中心组合旋转实验设计，利用53组实验筛选出它们的最优使用浓度分别为孕酮5．5mg／L、抗氧化试剂250μL/L、乙醇胺2．1mL／L、维生素B12 2．86mg／L、维生素B6 44μg/L和脂肪酸复合剂215μL/L。结论：经2种实验设计方法优化，确定了Marc-145细胞无血清培养液组分及最适浓度，为无血清悬浮培养Marc-145细胞及增殖猪繁殖及呼吸综合征病毒提供了实验基础。

Optimization of Serum Free Media for Marc-145 Cells Suspension Culture

Objective： To optimize the components of serum free media for Marc-145 ceils suspension culture. Methods： Plackett-Burman design and central composite rotatable design were adopted to optimize the different components in serum free media. Results： In the first step, according to Plackett-Burman design, 20 experimental groups were designed to evaluate the effects of 14 factors to the specific growth rate of Marc-145 cells in suspen- sion culture. By regression analysis, progesterone, anti-oxidant reagent, ethanolamine, vitamin Br, vitamin B~ and fatty acid reagent were found to have significant effects on the specific growth rate of Marc-145 cells. In the sec- ond step, central composite rotatable design was performed in 53 experimental groups to optimize the above criti- cal factors and to find out the optimal concentrations between these factors. Based on these results, the optimal concentrations were determined as 5.5 mg/L progesterone, 250μL/L anti-oxidant reagent, 2.1 mL/L ethanolamine, 2.86 mg/L vitamin B12, 44 μg/L vitamin B6 and 215 μL/L fatty acid reagent by solving the validated fitting qua- dratic regression model equation. Conclusion： The serum free media was screened out preliminarily through two steps optimization. This study provided the experimental data for Marc-145 cells suspension culture and porcine re- productive and respiratory syndrome virus（PRRSV） propagation in serum free media.