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A kind of full suspension mdck cell and the method utilizing full suspension mdck cell to cultivate swine influenza virus

A swine influenza virus and cell culture technology, applied in the field of fully suspended MDCK cells and the use of fully suspended MDCK cells to cultivate swine influenza virus, can solve problems such as virus mutation, long production cycle, and inability to respond to influenza pandemics in a timely and effective manner

Active Publication Date: 2020-12-04
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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  • A kind of full suspension mdck cell and the method utilizing full suspension mdck cell to cultivate swine influenza virus
  • A kind of full suspension mdck cell and the method utilizing full suspension mdck cell to cultivate swine influenza virus
  • A kind of full suspension mdck cell and the method utilizing full suspension mdck cell to cultivate swine influenza virus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Screening of MDCK Monoclonal Cell Strain and Swine Influenza Vaccine Strain

[0049] 1.1 Establishment of MDCK monoclonal cell bank

[0050] The adherent MDCK cells were subcloned three times by 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-E8 14 E8-D12-D3 2 D11-E3-C6 15 H7-F7-D9 3 D11-E3-D4 16 E8-D12-C3 4 E8-D12-A2 17 F7-G3-E10 5 D11-E6-A3 18 F7-G8-E2 6 D11-C3-C8 19 H7-F7-F9 7 D11-F11-D11 20 E8-D12-D5 8 D11-C3-E9 21 D11-E6-A4 9 H7-F7-G8 22 E8-D12-D2 10 D11-E6-D6 23 G10-B11-D5 11 D11-E6-E3 24 F7-G3-E8 12 D11-E6-C5 25 E11-D2-B11 13 H7-F7-E11

[0053] 1.2 Screening of SI vaccine strains

[0054] The first step: the preparation of hyperimmune serum. Use SPF...

Embodiment 2

[0062] Serum-free Suspension Acclimatization of MDCK Monoclonal Cell Line and Optimization of Virus Culture Conditions

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

[0064] normal culture at 25cm 2 Adherent MDCK cells in cell culture flasks were expanded to 75cm 2 cell culture flasks. Digest the adherent cells, count them, and press 9.0×10 5 cells / mL were transferred to a 125 mL cell shaker flask, and the culture volume was 25 mL. Set 80rpm, 100rpm, 120rpm, 130rpm, 135rpm, 140rpm, 145rpm, 150rpm to monitor cell viability at different speeds. The rotation speed corresponding to the highest cell viability was determined as the optimal rotation speed for cell suspension acclimatization, the results are shown in image 3 . from image 3 It can be concluded that, compared with other rotating speeds, when the rotating speed is 140rpm, the cell viability is the highest, which is 98.9%.

[0065] 2.2 Serum-free suspension a...

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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 invention belongs to the technical field of medicine or veterinary medicine, and in particular relates to a full-suspension MDCK cell and a method for cultivating swine influenza virus by using the full-suspension MDCK cell. Background technique [0002] Swine influenza (SI) is an acute, febrile and highly contagious respiratory infectious disease of pigs caused by swine influenza virus (SIV). Death cases are rare, but the mixed infection of other pathogens due to SIV infection often complicates and aggravates the disease, seriously endangers the health of pig herds, and causes huge economic losses to the farms. As an important zoonotic pathogen, SIV can be transmitted between humans and pigs, causing public health impacts. Influenza A virus poses a great threat to human beings. The "Spanish flu", "Asian flu", and "Hong Kong flu" that broke out in the last century caused tens of millions of deaths and caused great harm to human health and the econ...

Claims

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Application Information

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Patent Type & Authority Patents(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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