Madin-Darby canine kidney cell sialic acid receptor modulation induced by culture medium conditions: Implications for the isolation of influenza A virus

Influenza and Other Respiratory Viruses
Sarah W NelsonAndrew S Bowman

Abstract

The influenza A virus (IAV) binds to α-2,3- and α-2,6-linked sialic acid (SA) receptors expressed by Madin-Darby canine kidney (MDCK) cells. The receptor distribution may therefore be important in regulating IAV propagation. Serum-free medium (SFM) avoids variability in conventional culture medium containing fetal bovine serum (FBS), which can have variable composition and may contain endotoxins. However, little is known about the distribution of SA receptors on cells maintained in SFM. We assessed the influence of culture media on MDCK cell SA receptor distribution along with the effect of SA receptor distribution on IAV recovery. We hypothesized that SFM would increase the proportion of α-2,6-linked SA receptors present and alter isolate recovery. Madin-Darby canine kidney cells were cultured in medium containing FBS and two SFMs. Cell surface distribution of α-2,6- and α-2,3-linked receptors was determined using flow cytometry. Recovery of swine- and avian-lineage IAVs from MDCK cells maintained in each medium was quantified as TCID50 . Madin-Darby canine kidney cells cultured in UltraMDCK SFM expressed both SA receptors and supported the growth of both swine- and avian-lineage IAVs. Cells maintained in other medium inconsis...Continue Reading

References

Oct 24, 2001·Biochemistry·C L JacobsC R Bertozzi
Nov 15, 2008·Methods in Molecular Biology·Brett D Lindenbach
Feb 21, 2009·Vaccine·Nicole M Bouvier, Peter Palese
Apr 15, 2010·ALTEX·Daniel BrunnerGerhard Gstraunthaler
Jun 26, 2012·Journal of Virological Methods·Tina LombardoMaura Ferrari
Jun 29, 2012·The Journal of General Virology·Zhi SunYing Fang
Dec 18, 2012·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Andrew S BowmanRichard D Slemons
Feb 10, 2017·Transboundary and Emerging Diseases·J I Sánchez-BetancourtR A Segura-Velázquez

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Methods Mentioned

BETA
flow cytometry
protein assay
light microscopy

Software Mentioned

Stata

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