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犬肾传代细胞系 Madin-Darby 在无血清培养基中的悬浮培养适应及其与 Vero 和 BHK21 细胞系生产禽流感病毒能力的比较。

Adaptation of a Madin-Darby canine kidney cell line to suspension growth in serum-free media and comparison of its ability to produce avian influenza virus to Vero and BHK21 cell lines.

机构信息

Central Veterinary Institute of Wageningen UR (CVI), Lelystad, The Netherlands.

出版信息

J Virol Methods. 2011 Jan;171(1):53-60. doi: 10.1016/j.jviromet.2010.09.029. Epub 2010 Oct 7.

Abstract

Madin-Darby canine kidney (MDCK) cells are currently considered for influenza vaccine manufacturing. A drawback of these cells is their anchorage dependent growth, which greatly complicates process scale-up. In this paper a novel MDCK cell line (MDCK-SFS) is described that grows efficiently in suspension and retained high expression levels of both α-2,6 and α-2,3 sialic acid receptors, which bind preferably to human and avian influenza viruses, respectively. The production of avian influenza virus by BHK21, Vero and MDCK-SFS cell lines was compared. Although BHK21 cells consisted of two populations, one of which lacks the α-2,3 receptor, they supported the replication of two influenza strains to high titres. However, BHK21 cells are generally not applicable for influenza production since they supported the replication of six further strains poorly. MDCK-SFS cells yielded the highest infectious virus titres and virus genome equivalent concentration for five of the eight influenza strains analyzed and the highest hemagglutination activity for all eight virus strains. Taken together with their suitability for suspension growth this makes the MDCK-SFS cell line potentially useful for large scale influenza virus production.

摘要

Madin-Darby 犬肾 (MDCK) 细胞目前被认为可用于流感疫苗的生产。这些细胞的一个缺点是其锚定依赖性生长,这极大地增加了工艺放大的复杂性。本文描述了一种新型的 MDCK 细胞系(MDCK-SFS),该细胞系在悬浮培养中高效生长,并保持高水平的α-2,6 和 α-2,3 唾液酸受体表达,分别与人类和禽流感病毒结合。比较了 BHK21、Vero 和 MDCK-SFS 细胞系生产禽流感病毒的情况。尽管 BHK21 细胞由两个群体组成,其中一个缺乏α-2,3 受体,但它们支持两种流感株的高滴度复制。然而,BHK21 细胞通常不适用于流感生产,因为它们对另外六种流感株的复制支持不佳。MDCK-SFS 细胞系对分析的八种流感株中的五种产生了最高的感染性病毒滴度和病毒基因组等效浓度,对所有八种病毒株产生了最高的血凝活性。加上它们适合悬浮培养,这使得 MDCK-SFS 细胞系有可能用于大规模流感病毒生产。

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