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[Optimization of culture condition for cultivation of influenza virus A on MDCK cell]
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.
Insights
Optimizing influenza virus A cultivation in MDCK cells revealed specific conditions for swine-origin H1N1, seasonal H1N1, and seasonal H3N2. Swine-origin H1N1 showed lower separation efficiency compared to seasonal strains.
Area of Science:
- Virology
- Cell Biology
- Infectious Diseases
Background:
- Influenza A virus poses a significant public health threat.
- MDCK cells are a standard model for influenza virus propagation.
- Optimizing cultivation conditions is crucial for research and diagnostics.
Purpose of the Study:
- To determine optimal cultivation parameters for Influenza A virus subtypes on MDCK cells.
- To enhance the separation rate and diagnostic efficacy of influenza virus.
- To compare the cultivation characteristics of swine-origin H1N1, seasonal H1N1, and seasonal H3N2.
Main Methods:
- Cultivation of three Influenza A virus strains (swine-origin H1N1, seasonal H1N1, seasonal H3N2) on MDCK cells.
- Systematic variation of inoculum size, TPCK-Trypsin concentration, and MDCK cell passage generations.
- Evaluation of virus growth and infectivity through hemagglutination inhibition (HI) assays.
Main Results:
- Optimal inoculum sizes were 100 µl/cm² for swine-origin H1N1 and 75 µl/cm² for seasonal H1N1 and H3N2.
- Optimal TPCK-Trypsin concentrations were 2.5 µg/ml for swine-origin H1N1 and 2.0 µg/ml for seasonal strains.
- Swine-origin H1N1 showed higher susceptibility in earlier MDCK cell generations but lower HI titers post-cultivation compared to seasonal strains.
Conclusions:
- Swine-origin H1N1 exhibited inferior separation efficiency in MDCK cells compared to seasonal H1N1 and H3N2.
- Optimal cultivation conditions for seasonal H1N1 and H3N2 were found to be similar.
- Further optimization may be needed for efficient swine-origin H1N1 isolation and study.
