Application of a serum-free medium for the growth of vero cells and the production of reovirus
Application of a serum-free medium for the growth of vero cells and the production of reovirus
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DOI：
10.1021/bp000110
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发表时间：
2000-09-01
影响因子：
2.9
通讯作者：
Coombs, K
中科院分区：
文献类型：
--
作者：
Butler, M;Burgener, A;Coombs, K
两株呼肠孤病毒(血清1型Lang/TIL和血清3型Dearing/T3d)在无血清M-VSFM的静止和搅拌培养的Vero细胞中增殖。固体微载体(Cytodex-1)用于支持细胞在正常倍增时间为25h的搅拌培养中生长，在微载体培养中，接种量为2×10(5)个/毫升，4d后细胞产量可达1×10(6)个/毫升。生长曲线和细胞产量与添加血清的培养没有显著差异。在150h的培养时间内，病毒滴度增加了3-4个数量级，在M-VSFM中，两株呼肠孤病毒在静止培养上的最大病毒滴度都达到了>；1×10(9)pfu/mL。M-VSFM还支持在微载体培养中获得高产量的病毒。M-VSFM的比产和最终病毒产量均高于血清补充组。病毒的高产量表明，这是一个适合生产呼肠孤病毒的系统，作为人类治疗用的溶瘤剂。
Two strains of reovirus (serotype 1 Lang/TIL and serotype 3 Dearing/T3D) were propagated in Vero cells grown in stationary or agitated cultures in a serum-free medium, M-VSFM. Solid microcarriers (Cytodex-1) were used to support cell growth in agitated cultures with a normal doubling time of 25 h. Cell yields of 1 x 10(6) cells/mL were obtained from an inoculum of 2 x 10(5) cells/mL in 4 days in microcarrier cultures. The growth profile and cell yield was not significantly different from serum-supplemented cultures. The virus titer increased by 3-4 orders of magnitude over a culture period of 150 h. The maximum virus titer in stationary cultures reached >1 x 10(9) pfu/mL for both strains of reovirus in M-VSFM. M-VSFM also supported high viral yields in microcarrier cultures. Both the specific productivity and final viral yield was higher in M-VSFM than serum-supplemented cultures. The high viral productivity suggests that this is a suitable system for the production of reovirus as an oncolytic agent for human therapeutic use.