A kind of vero E6 cell line adapted to full suspension culture and application thereof

Abstract

The invention discloses a vero E6 cell strain adapting to full-suspension culture, the vero E6 cell strain is named as vero E6-s, and is preserved in China Center for Type Culture Collection, the preservation number is CCTCC2017101, the vero E6 cell strain is classified and named as Vero cell suspension adaptive strain VeroE6-S, and the preservation date is June 29, 2017. Compared with the prior art, the vero E6 cell strain adapting to the full-suspension culture has high degree of automation for culture of vaccine viruses, can be suspended and cultured in a serum-free or low-serum medium without carrier intervention, and solves large-scale industrialization requirements of virus cultivation, a large-scale production method meeting GMP production technology requirements can be developed, and the large-scale production method has a good prospect of industrialization.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a vero E6 cell strain adapted to full suspension culture and application thereof. Background technique [0002] Vero-E6 cells are the African green monkey kidney cell line, which belongs to epithelial cells and is derived from the African green monkey kidney cells. It is a clone of the mother cell Vero 76, grows in an anchorage-dependent manner, can be continuously subcultured, and has no tumorigenicity. Suspension-adapted strains are obtained after several times of suspension acclimatization of cell lines, which can grow in suspension without relying on the carrier. The cells are particularly sensitive to porcine epidemic diarrhea virus, rabies virus, porcine pseudorabies virus, canine distemper virus, canine adenovirus, and canine parainfluenza virus, and are currently widely used in the production of vaccines for these viral diseases. [0003] Due to the attachment-de...

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Problems solved by technology

[0003] Due to the attachment-dependent growth of the original Vero-E6 cells, the production of these viral disease vaccines mainly adopts the traditional spinner bottle process. The production process of vaccine antigen preparation is labor-intensive and the probability of contamination is high, and the quality of the vaccine is unstable. The difference is large. This traditional process is gradually replaced by automated production in today's large-scale industrial production needs.

Bioreactor cell suspension culture has become the main research force of the current vaccine industry automation production process. Because of the difficulty of cell domestication and long research and development cycle, many enterprises choose carrier-dependent cell suspension culture. A method for preparing porcine epidemic diarrhea virus in a torrent bioreactor" and the patent publication number is CN 101804203 A "A method for large-scale production of pseudorabies virus vaccine", and the patent publication number is CN 102038946A "Industrial production of pseudorabies virus vaccine using bioreactor The method of rabies vaccine" and the patent publication number are CN 104587460 A "mink viral enteritis, canine distemper dual live vaccine and its preparation method and application"; although this kind of microcarrier and paper carrier suspension culture is more traditional The degree of automation is high, but cell culture is limited by carrier factors and cannot achieve optimal growth. Compared with cell suspension culture, carrier-dependent culture increases the production process and increases production costs. The degree of scale-up is limited, and it is only the initial stage of cell suspension culture.

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