响应面法优化MDCK-siat7e细胞培养工艺

摘要

Objective To optimize MDCK-siat7e cell culture conditions by response surface methodology (RSM).Methods Initial ammonia concentration,lactic acid concentration,osmotic pressure and viable cell density (VCD) of MDCK-siat7e cell culture were used as factors.Cell specific growth rate,cell viability,lactic acid and ammonia accumulation ratios of MDCK-siat7e cell culture at 72 h were set as response values.The full factorial design and RSM were used to study the effect of these factors on MDCK-siat7e ceil culture.Results During MDCK-siat7e cells culture,initial VCD was a significant factor effecting on specific growth rate (t =3.43,P ＜0.05);initial ammonia and lactic acid concentrations were significant factors effecting on cell viability (t =4.32,P ＜0.05;t =2.86,P ＜0.05);initial lactic acid concentration,osmotic pressure and VCD were significant factors effecting on lactic acid accumulation ratio (t =8.92,P ＜0.05;t =4.11,P ＜0.05;t =2.66,P ＜0.05),and initial lactic acid concentration played a major role.Initial ammonia concentration was a significant factor effecting on ammonia accumulation ratio (t =2.63,P ＜0.05).Conclusion Using RSM to optimize MDCK-siat7e cell culture process can guide MDCK-siat7e cell culture condition control.%目的 以响应面法优化MDCK-siat7e细胞培养条件.方法 以MDCK-siat7e细胞培养的起始氨浓度、乳酸浓度、渗透压和活细胞密度(viable cell density,VCD)为因子,以MDCK-siat7e细胞培养的72 h细胞比生长率、细胞活率、乳酸比生成率和氨比生成率为响应值,采用全因子实验设计和响应面分析来研究这些因子对MDCK-siat7e细胞培养的影响.结果 在MDCK-siat7e细胞培养过程中,对72 h细胞比生长率有显著影响的因子是起始VCD(t=3.43,P＜0.05);对72 h细胞活率有显著影响的因子是起始乳酸浓度(t=4.32,P＜0.05)和氨浓度(t=2.86,P＜0.05);对72 h乳酸比生成率有显著影响的因子是起始乳酸浓度(t=8.92,P＜0.05)、渗透压(t=4.11,P＜0.05)和VCD(t=2.66,P＜0.05),其中乳酸浓度起主要作用;对72 h氨比生成率有显著影响的因子是起始氨浓度(t=2.63,P＜0.05).结论 用响应面法优化MDCK-siat7e细胞培养工艺可对该细胞培养过程中的条件控制起指导作用.