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基于微载体的细胞培养工艺生产牛呼吸道合胞体病毒疫苗。

A microcarrier-based cell culture process for the production of a bovine respiratory syncytial virus vaccine.

机构信息

MFM Laboratories Ltd., 4 Warner Dr., Springwood Industrial Estate, Braintree, Essex, CM7 2YW, UK.

出版信息

Cytotechnology. 1999 Mar;29(2):135-49. doi: 10.1023/A:1008022828736.

Abstract

Veterinary viral vaccines generally comprise either attenuated or chemically inactivated viruses which have been propagated on mammalian cell substrates or specific pathogen free (SPF) eggs. New generation vaccines include chemically inactivated virally-infected whole cell vaccines. The NM57 cell line is a bovine nasal turbinate persistently infected (non-lytic infection) with a strain of the respiratory syncytial virus (RSV). The potential of microcarrier technology for the cultivation in bioreactors of this anchorage dependent cell line for RSV vaccine production has been investigated. Both Cytodex 3 and Cultispher S microcarriers proved most suitable from a selection of microcarriers as growth substrates for this NM57 cell line. Maximum cell densities of 4.12×105 cells ml-1and 5.52×105 cells ml-1 respectively were obtained using Cytodex 3 (3 g l-1) and and Cultispher S (1 g l-1) in 5 l bioreactor cultures. The fact that cell growth was less sensitive to agitation rate when cultured on Cultispher S microcarriers, and that cells were efficiently harvested from this microcarrier by an enzymatic method, suggested Cultispher S is suitable for further evaluation at larger bioreactor scales (>5 l) than that described here.

摘要

兽医病毒疫苗通常包括减毒或化学灭活的病毒,这些病毒已在哺乳动物细胞基质或无特定病原体(SPF)卵上繁殖。新一代疫苗包括化学灭活的感染性全细胞疫苗。NM57 细胞系是一种牛鼻甲骨持续感染(非裂解感染)的呼吸道合胞病毒(RSV)株。已经研究了微载体技术在生物反应器中用于 RSV 疫苗生产的这种锚定依赖性细胞系培养的潜力。在微载体的选择中,Cytodex 3 和 Cultispher S 微载体均被证明是最适合作为 NM57 细胞系生长基质的微载体。在 5 l 生物反应器培养中,使用 Cytodex 3(3 g l-1)和 Cultispher S(1 g l-1)分别获得了 4.12×105 个细胞 ml-1和 5.52×105 个细胞 ml-1的最大细胞密度。当在 Cultispher S 微载体上培养时,细胞生长对搅拌速度的敏感性较低,并且可以通过酶法有效地从该微载体中收获细胞,这表明 Cultispher S 适合在比此处描述的更大的生物反应器规模(>5 l)进行进一步评估。

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Cytotechnology. 1995 Jan;18(1-2):51-6. doi: 10.1007/BF00744319.
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Optimization of physical parameters for cell attachment and growth on macroporous microcarriers.用于细胞在大孔微载体上附着和生长的物理参数优化。
Biotechnol Bioeng. 1996 Jun 20;50(6):627-35. doi: 10.1002/(SICI)1097-0290(19960620)50:6<627::AID-BIT3>3.0.CO;2-M.
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Growth and death in overagitated microcarrier cell cultures.
Biotechnol Bioeng. 1989 Feb 5;33(6):731-44. doi: 10.1002/bit.260330611.

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