Full-suspension MDCK cell and method for culturing swine influenza virus by using full-suspension MDCK cell

A swine influenza virus and cell culture technology, which is applied in the field of fully suspended MDCK cells and the use of fully suspended MDCK cells to cultivate swine influenza virus, can solve the problems of inability to timely and effectively respond to influenza pandemics, cumbersome chicken embryo production operations, and body allergies.

Active Publication Date: 2020-07-24
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current production medium of influenza vaccines is mainly chicken embryos. This method has many defects, such as too long production cycle, insufficient supply of chicken embryos when the epidemic occurs, easy to cause body allergies and virus mutations, etc., and the production of chicken embryos is cumbersome. Inability to respond to a pandemic in a timely and effective manner

Method used

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  • Full-suspension MDCK cell and method for culturing swine influenza virus by using full-suspension MDCK cell
  • Full-suspension MDCK cell and method for culturing swine influenza virus by using full-suspension MDCK cell
  • Full-suspension MDCK cell and method for culturing swine influenza virus by using full-suspension MDCK cell

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Screening of MDCK monoclonal cell strain and swine flu vaccine strain

[0049] 1.1 Establishment of MDCK monoclonal cell bank

[0050] The adherent MDCK cells were subcloned 3 times using the limiting dilution method to obtain 25 MDCK monoclonal cell lines. The results are shown in Table 1.

[0051] Table 1 MDCK monoclonal cell bank

[0052] Cell line number Cell line name Cell line number Cell line name 1 D11-C3-E814 E8-D12-D3 2 D11-E3-C615 H7-F7-D9 3 D11-E3-D416 E8-D12-C3 4 E8-D12-A217 F7-G3-E10 5 D11-E6-A318 F7-G8-E2 6 D11-C3-C819 H7-F7-F9 7 D11-F11-D1120 E8-D12-D5 8 D11-C3-E921 D11-E6-A4 9 H7-F7-G822 E8-D12-D2 10 D11-E6-D623 G10-B11-D5 11 D11-E6-E324 F7-G3-E8 12 D11-E6-C525 E11-D2-B11 13 H7-F7-E11

[0053] 1.2 Screening of SI vaccine strains

[0054] The first step: preparation of high immunity serum. Use SPF chicken embryos to multiply SIV of different genotypes, harvest the allantoic fluid and inactivate it with formaldehyde, after preparing the vaccine, immunize SPF ...

Embodiment 2

[0062] Serum-free suspension acclimation of MDCK monoclonal cell line and optimization of virus culture conditions

[0063] 2.1 Establish the speed of suspension acclimation of MDCK monoclonal cell line.

[0064] Culture normal at 25cm 2 The adherent MDCK cells in the cell culture flask are expanded to 75cm 2 Cell culture flask. Digest the adherent cells, and after counting, press 9.0×10 5 One cell / mL was connected to a 125mL cell shaker flask with a culture volume of 25mL. Set 80rpm, 100rpm, 120rpm, 130rpm, 135rpm, 140rpm, 145rpm, 150rpm to monitor the cell viability at different speeds. Determine the speed corresponding to the highest cell viability as the best speed for cell suspension acclimation. image 3 . From image 3 It can be concluded that, compared with other speeds, when the speed is 140 rpm, the cell viability is the highest, which is 98.9%.

[0065] 2.2 Serum-free suspension domestication of MDCK monoclonal cell line

[0066] Culture normal at 25cm 2 The adherent cel...

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Abstract

The invention provides a full-suspension MDCK cell and a method for culturing swine influenza virus by using the full-suspension MDCK cell, and belongs to the technical field of medicine or veterinarymedicine. The full-suspension MDCK cell is a full-suspension MDCK cell D11-F11-D1. The full-suspension MDCK cell provided by the invention can proliferate swine influenza virus with high titer, provides a good culture medium for the research and development and production of SI vaccines, and lays a foundation for developing a cell culture process for the production of SI vaccines.

Description

Technical field [0001] The present invention belongs to the technical field of medicine or veterinary medicine, and in particular relates to a method for culturing swine influenza virus by using the fully suspended MDCK cells and the fully suspended MDCK cells. Background technique [0002] Swine influenza (SI) is an acute, febrile and highly contact respiratory infectious disease of swine caused by swine influenza virus (SIV). The clinical symptoms are usually mild respiratory diseases, with high incidence and rapid recovery. Deaths are rare, but mixed infections of other pathogens due to SIV infection often complicate and aggravate the disease, seriously endanger the health of the pig herd, and cause huge economic losses to the farm. As an important pathogen of zoonotic diseases, SIV can be transmitted between humans and pigs, and affect public health. The influenza A virus poses a great threat to humans. The pandemics such as "Spanish influenza", "Asian influenza" and "Hong K...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N7/00C12R1/93
CPCC12N5/0686C12N7/00C12N2509/00C12N2760/16151
Inventor 乔传玲陈化兰杨焕良陈艳
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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