CAJ | 학술논문

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 B6、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5.5 mg/L、抗氧化试剂250μL/L、乙醇胺2.1 mL/L、维生素 B122.86 mg/L、维生素 B644μg/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.
목적:대 Marc-145세포무혈청배양액조분급농도진행우화사선.방법:채용 Plackett-Burman 설계화중심조합선전설계대불동적영양성분진행우화사선.결과:경 Plackett-Burman 설계,이용20조실험대14충불동영양성분진행고찰,발현잉동、항양화시제、을순알、유생소 B6、유생소 B12화지방산복합제대 Marc-145세포무혈청현부생장적비생장속솔유현저영향;침대해6충인소진행중심조합선전실험설계,이용53조실험사선출타문적최우사용농도분별위잉동5.5 mg/L、항양화시제250μL/L、을순알2.1 mL/L、유생소 B122.86 mg/L、유생소 B644μg/L 화지방산복합제215μL/L.결론:경2충실험설계방법우화,학정료 Marc-145세포무혈청배양액조분급최괄농도,위무혈청현부배양 Marc-145세포급증식저번식급호흡종합정병독제공료실험기출.
Objective: To optimize the components of serum free media for Marc-145 cells 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 B6, vitamin B12 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, 250 μL/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.