Evaluation of the serum-free medium MDSS2 for the production of poliovirus on Vero cells in bioreactors
Evaluation of the serum-free medium MDSS2 for the production of poliovirus on Vero cells in bioreactors
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DOI：
10.1023/a:1007999313566
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发表时间：
1997-01-01
期刊：
影响因子：
2.2
通讯作者：
Crainic, R
中科院分区：
文献类型：
--
作者：
Merten, OW;Wu, R;Crainic, R
无血清培养基MDSS 2（Merten等人，1994）用于培养Vero细胞以及用于在静态和灌注微载体培养物中产生脊髓灰质炎病毒（Sabin 1型）。在分别为0.0120/h和0.0106/h的略有不同的生长速率下，含血清（SCM）和无血清（SFM）培养基中的静态培养物产生的滴度为10（6.75）和10（6.67）TCID50/50 μ l;表明比生产率为0.89和1.07 TCID 50/c。Vero细胞在6.25 g/ml的DEAE-葡聚糖微载体上的无血清生物反应器培养物。L产生的细胞密度为约1.5 × 10（6）C/ml。病毒感染后（感染复数（MOI）0.1-0.3），滴度约为6.3 × 10（8）TCID 50/ml，表明平均比生产率为7.1 TCID 50/c.h.。尽管这些值分别比经典的基于乙醇的生产方法高4倍和2倍（Montagnon等人，1981），一种参考培养物，其细胞生长在SCM中进行，仅病毒生产在SFM中进行，产生2 x 10（9）TCID/ml，平均比病毒生产率为18.9 TCID 50/c.h.。完全无血清和我们的参考工艺之间的差异主要是由于在SCM和SFM中生长的细胞的生理差异，以及由于病毒感染后消耗动力学的强烈改变，导致一种或几种必需培养基化合物（如葡萄糖和氨基酸）的限制。通过增加葡萄糖、谷氨酰胺、组氨酸和SH-氨基酸的残留浓度来避免这些限制，导致特异性病毒产生速率（约17.9 TCID 50/c.h.）与参考病毒生产工艺中发现的那些相当。脊髓灰质炎病毒（Sabin 1）生产的优化将根据培养基组成的修改进行描述。
The serum-free medium MDSS2 (Merten et al., 1994), was used for cultivating Vero cells as well as for producing poliovirus (Sabin type 1) in static and in perfused microcarrier cultures. At slightly different growth rates of 0.0120/h and 0.0106/h, respectively, static cultures in serum-containing (SCM) and serum-free (SFM) medium produced titers of 10(6.75) and 10(6.67) TCID50 per 50 mu l; signifying a specific productivity of 0.89 and 1.07 TCID50/c.Serum-free bioreactor cultures of Vero cells on DEAE-dextran microcarriers at 6.25 g/l produced cell densities of about 1.5 x 10(6)c/ml. After infection with virus (multiplicity of infection (MOI) 0.1-0.3) titers of about 6.3 x 10(8) TCID50/ml were obtained, signifying an average specific productivity of 7.1 TCID50/c.h. Although these values were 4 and 2 fold, respectively, higher than in classical resum-based production processes (Montagnon et al., 1981), a reference culture, for which cell growth was done in SCM and only virus production was done in SFM, produced 2 x 10(9) TCID/ml with an average specific virus production rate of 18.9 TCID50/c.h. The differences between the fully serum-free and our reference process were mainly due to physiological differences of cells grown in SCM and SFM and also due to strongly modified consumption kinetics after virus infection leading to limitations of one or several essential medium compounds, like glucose and amino acids. Avoiding these limitations by increasing the residual concentration of glucose, glutamine, histidine, and SH-amino acids, led to specific virus production rates (of about 17.9 TCID50/c.h.) comparable to those found in the reference virus production process. The optimisation of the production of the poliovirus (Sabin 1) will be described with respect to the modification of the medium composition.