Metabolism of MDCK cells during cell growth and influenza virus production in large-scale microcarrier culture.
Abstract: The production of equine influenza in Madin-Darby canine kidney (MDCK) cells in large-scale microcarrier culture is described with detailed on- and off-line analytical data during cell growth and virus replication. Metabolite concentration profiles for glucose, glutamine, lactate and ammonium are shown. Lactate and ammonium concentrations were always below inhibiting levels. Concentration profiles for essential and non-essential amino acids of the cell culture medium are discussed. During cell growth proline was released into the medium with a significant rate while two amino acids, serine and methionine were almost depleted. After infection, virus titer increased after a delay of 10-16 h whereas first changes in amino acid metabolism could be observed within 4h post-infection. Here, glutamate and aspartate increase correlated to virus release kinetics, indicating cell disruption and apoptosis. Starting with a moi of 0.025 resulted in a maximum virus yield of 2.4 log HA/100 microl at 44 h post-infection.
Publication Date: 2004-05-20 PubMed ID: 15149778DOI: 10.1016/j.vaccine.2003.11.041Google Scholar: Lookup
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- Journal Article
Summary
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The research article explores the metabolic activities of a specific type of cell, Madin-Darby canine kidney (MDCK) during its growth and while producing influenza virus in a large-scale microcarrier culture. The focus is on determining the concentration levels of certain metabolites and the behavior of essential and non-essential amino acids in the process.
Metabolite Concentration Profiles
- The research provides an extensive analytical look into the metabolite concentration profiles of glucose, glutamine, lactate, and ammonium during the growth and influenza virus replication in MDCK cells.
- According to the study, the concentrations of lactate and ammonium were always maintained below inhibiting levels. This suggests that the cells were kept in conditions that did not hinder their metabolic functions while they produced the virus.
Amino Acid Changes in Cell Culture Medium
- This study also delves into understanding the concentration profiles of essential and non-essential amino acids in the cell culture medium during this process.
- It was found that during cell growth, the amino acid proline was released into the medium at a significant rate, while two others, serine and methionine, were almost completely depleted. This implies that these three amino acids play key roles in the metabolism of MDCK cells during their growth.
Transition Post Infection
- Changes were also observed post infection. The virus titer displayed an increase after an initial lag phase of 10-16 hours following infection.
- Moreover, initial shifts in the amino acid metabolism were noted within just four hours. Glutamate and aspartate levels were found to rise in correlation with the kinetics of virus release, suggesting an increase in cell disruption and apoptosis.
Virus Yield
- The experiment began with a multiplicity of infection (moi) of 0.025. This low starting point led to a maximum virus yield of 2.4 log HA/100 microliters at 44 hours post-infection, signifying the efficiency and potential scale of virus production using MDCK cells in large-scale microcarrier cultures.
Cite This Article
APA
Genzel Y, Behrendt I, König S, Sann H, Reichl U.
(2004).
Metabolism of MDCK cells during cell growth and influenza virus production in large-scale microcarrier culture.
Vaccine, 22(17-18), 2202-2208.
https://doi.org/10.1016/j.vaccine.2003.11.041 Publication
Researcher Affiliations
- Max-Planck-Institute for Dynamics of Complex Technical Systems, Magdeburg, Sandtorstr. 1, 39106 Magdeburg, Germany. genzel@mpi-magdeburg.mpg.de
MeSH Terms
- Amino Acids / metabolism
- Animals
- Apoptosis
- Aspartic Acid / metabolism
- Cell Line
- Cell Proliferation
- Cells, Immobilized
- Dogs
- Glucose / metabolism
- Glutamine / metabolism
- Hemagglutinin Glycoproteins, Influenza Virus / analysis
- Influenza A virus / growth & development
- Kinetics
- Lactic Acid / metabolism
- Methionine / metabolism
- Proline / metabolism
- Quaternary Ammonium Compounds / metabolism
- Serine / metabolism
- Virus Cultivation / methods
- Virus Replication
Citations
This article has been cited 25 times.- Ramos JRC, Bissinger T, Genzel Y, Reichl U. Impact of Influenza A Virus Infection on Growth and Metabolism of Suspension MDCK Cells Using a Dynamic Model.. Metabolites 2022 Mar 12;12(3).
- Ren L, Zhang W, Zhang J, Zhang J, Zhang H, Zhu Y, Meng X, Yi Z, Wang R. Influenza A Virus (H1N1) Infection Induces Glycolysis to Facilitate Viral Replication.. Virol Sin 2021 Dec;36(6):1532-1542.
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