Heterogeneity of the MDCK cell line and its applicability for influenza virus research.

Single-cell clones have been established from the MDCK cell line, characterized for their morphology and evaluated for their suitability for influenza virus research. Three discrete cell morphotypes were identified using light microscopy. Besides morphological features, the cell types can be disting...

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Autores principales:	Vladimir Y Lugovtsev, Darya Melnyk, Jerry P Weir
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6857641d4a48410fa96500e12b0a6325
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spelling	oai:doaj.org-article:6857641d4a48410fa96500e12b0a63252021-11-18T08:55:17ZHeterogeneity of the MDCK cell line and its applicability for influenza virus research.1932-620310.1371/journal.pone.0075014https://doaj.org/article/6857641d4a48410fa96500e12b0a63252013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24058646/?tool=EBIhttps://doaj.org/toc/1932-6203Single-cell clones have been established from the MDCK cell line, characterized for their morphology and evaluated for their suitability for influenza virus research. Three discrete cell morphotypes were identified using light microscopy. Besides morphological features, the cell types can be distinguished by the level of expression of surface glycans recognized by peanut agglutinin (PNA). All clones were susceptible to infection by influenza viruses of different subtypes of influenza A virus (H1N1, H1N1pdm09, H3N2, H5N1) and influenza B virus, and all possessed on their surface terminally sialylated glycans with both types of glycosidic linkage (α2-3 and α2-6). The Type-1 cell lines were able to support a multicycle replication of influenza A and B viruses without help of an exogenous trypsin. In contrast, cell lines exhibiting Type-2 morphology were unable to support multicycle replication of influenza A viruses without trypsin supplementation. Western blot analysis of the hemagglutinin of H1N1 strains demonstrated that Type-2 cells were deficient in production of proteolytically activated hemagglutinin (no cleavage between HA1/HA2 was observed). HA1/HA2 cleavage of influenza B viruses in the Type-2 cells was also significantly impaired, but not completely abrogated, producing sufficient amount of activated HA to support efficient virus replication without trypsin. In contrast, all clones of Type-1 cells were able to produce proteolytically activated hemagglutinin of influenza A and B viruses. However, the growth kinetics and plaque size of influenza A viruses varied significantly in different clones. Influenza B virus also showed different plaque size, with the biggest plaque formation in the Type-2 cells, although the growth kinetics and peak infectivity titers were similar in all clones. Taken together, the study demonstrates that the population of original MDCK cells is represented by various types of cells that differ in their capacities to support replication of influenza A and B viruses.Vladimir Y LugovtsevDarya MelnykJerry P WeirPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 9, p e75014 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Vladimir Y Lugovtsev Darya Melnyk Jerry P Weir Heterogeneity of the MDCK cell line and its applicability for influenza virus research.
description	Single-cell clones have been established from the MDCK cell line, characterized for their morphology and evaluated for their suitability for influenza virus research. Three discrete cell morphotypes were identified using light microscopy. Besides morphological features, the cell types can be distinguished by the level of expression of surface glycans recognized by peanut agglutinin (PNA). All clones were susceptible to infection by influenza viruses of different subtypes of influenza A virus (H1N1, H1N1pdm09, H3N2, H5N1) and influenza B virus, and all possessed on their surface terminally sialylated glycans with both types of glycosidic linkage (α2-3 and α2-6). The Type-1 cell lines were able to support a multicycle replication of influenza A and B viruses without help of an exogenous trypsin. In contrast, cell lines exhibiting Type-2 morphology were unable to support multicycle replication of influenza A viruses without trypsin supplementation. Western blot analysis of the hemagglutinin of H1N1 strains demonstrated that Type-2 cells were deficient in production of proteolytically activated hemagglutinin (no cleavage between HA1/HA2 was observed). HA1/HA2 cleavage of influenza B viruses in the Type-2 cells was also significantly impaired, but not completely abrogated, producing sufficient amount of activated HA to support efficient virus replication without trypsin. In contrast, all clones of Type-1 cells were able to produce proteolytically activated hemagglutinin of influenza A and B viruses. However, the growth kinetics and plaque size of influenza A viruses varied significantly in different clones. Influenza B virus also showed different plaque size, with the biggest plaque formation in the Type-2 cells, although the growth kinetics and peak infectivity titers were similar in all clones. Taken together, the study demonstrates that the population of original MDCK cells is represented by various types of cells that differ in their capacities to support replication of influenza A and B viruses.
format	article
author	Vladimir Y Lugovtsev Darya Melnyk Jerry P Weir
author_facet	Vladimir Y Lugovtsev Darya Melnyk Jerry P Weir
author_sort	Vladimir Y Lugovtsev
title	Heterogeneity of the MDCK cell line and its applicability for influenza virus research.
title_short	Heterogeneity of the MDCK cell line and its applicability for influenza virus research.
title_full	Heterogeneity of the MDCK cell line and its applicability for influenza virus research.
title_fullStr	Heterogeneity of the MDCK cell line and its applicability for influenza virus research.
title_full_unstemmed	Heterogeneity of the MDCK cell line and its applicability for influenza virus research.
title_sort	heterogeneity of the mdck cell line and its applicability for influenza virus research.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/6857641d4a48410fa96500e12b0a6325
work_keys_str_mv	AT vladimirylugovtsev heterogeneityofthemdckcelllineanditsapplicabilityforinfluenzavirusresearch AT daryamelnyk heterogeneityofthemdckcelllineanditsapplicabilityforinfluenzavirusresearch AT jerrypweir heterogeneityofthemdckcelllineanditsapplicabilityforinfluenzavirusresearch
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Heterogeneity of the MDCK cell line and its applicability for influenza virus research.

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