[Optimization of culture condition for cultivation of influenza virus A on MDCK cell]

Wei Sheng Yan Jiu. 2014 Mar;43(2):210-2, 218.
[Article in Chinese]

Abstract

Objective: To optimize the cultivation condition of influenza virus A on MDCK cells and to improve the separation rate and the inspected effect of influenza virus.

Methods: The virus specimens which came from three kinds of the influenza virus A, swine-origin H1N1, seasonal H1N1 and seasonal H3N2, were cultivated by MDCK cells. It will make the most suitable condition by means of comparing the different inoculum size (25, 50, 75, 100, 125 and 150 microl/cm2), the different concentration of TPCK-Trypsin (1.0, 1.5, 2.0, 2.5, 3.0 and 3.5 microg/ml) as an addition and the different susceptibility of cell generations for serial passage culture of four generations of these three influenza virus.

Results: Throughout the comparison the three kinds of the influenza virus, swine-origin H1N1, seasonal H1N1 and seasonal H3N2, which had the most suitable inoculum size 100,75, 75 microl/cm2 and the most suitable concentration of TPCK-Trypsin as an addition 2.5, 2.0, 2.0 microg/ml and the most suitable generations of cell 1, 2, 2 respectively. Earlier generations of MDCK cells of the swine-origin H1N1 were more susceptible than the other influenza virus A. The hemagglutination inhibition (HI) of swine-origin H1N1 after cultivated was lower than the other of seasonal H1N1 and seasonal H3N2.

Conclusion: The separating effect of the swine-origin H1N1 virus in the MDCK cell was inferior to the seasonal H1N1 and seasonal H3N2. The culture condition of seasonal H1N1 and seasonal H3N2 were roughly similar.

MeSH terms

  • Alphainfluenzavirus
  • Animals
  • Cell Culture Techniques
  • Dogs
  • Influenza A Virus, H1N1 Subtype*
  • Influenza A Virus, H3N2 Subtype*
  • Madin Darby Canine Kidney Cells
  • Swine