Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses

Abstract A stable culture of primary porcine enterocytes is necessary to study porcine enteric virus replication characteristics. Because the direct cultivation of primary porcine enterocytes is difficult, alternatives have to be considered. As subepithelial myofibroblasts secrete extracellular matr...

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Autores principales:	Tingting Cui, Sebastiaan Theuns, Lowiese M. B. Desmarets, Jiexiong Xie, Gaëtan M. A. De Gryse, Bo Yang, Wim Van den Broeck, Hans J. Nauwynck
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Porcine Epidemic Diarrhea Virus (PEDV) Transmissible Gastroenteritis Virus (TGEV) PEDV CV777 Rotavirus Strains Milan Strain Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dd10812196a04031b5d536d14c9aa44f
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spelling	oai:doaj.org-article:dd10812196a04031b5d536d14c9aa44f2021-12-02T15:08:42ZEstablishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses10.1038/s41598-018-33305-12045-2322https://doaj.org/article/dd10812196a04031b5d536d14c9aa44f2018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-33305-1https://doaj.org/toc/2045-2322Abstract A stable culture of primary porcine enterocytes is necessary to study porcine enteric virus replication characteristics. Because the direct cultivation of primary porcine enterocytes is difficult, alternatives have to be considered. As subepithelial myofibroblasts secrete extracellular matrix and growth factors contributing to the attachment, proliferation and differentiation of epithelial cells, co-cultures of primary porcine enterocytes (ileocytes and colonocytes) with myofibroblasts were developed and evaluated for their susceptibility to enteric viruses. First, it was demonstrated that the co-cultured ileocytes and colonocytes were susceptible to an archival rotavirus strain RVA/pig-tc/BEL/RV277/1977/G1P[7] and different other rotavirus genotypes (fecal samples containing G5P[7], G5P[13], G9P[23], G4P[6]). Next, the TGEV Purdue strain infected both ileocytes and colonocytes whereas the Miller strain only infected ileocytes. Last, the PEDV CV777 Vero adapted and non-adapted (fecal suspension) strains could infect co-cultured ileocytes but not colonocytes. The infectivity of the CV777 Vero adapted strain was higher when the cells were cultured without fetal bovine serum and the CV777 fecal suspension only infected the ileocytes cultured without fetal bovine serum. In conclusion, a novel co-culture of porcine enterocytes with myofibroblasts was established, which can be used for the investigation of the replication of enteric viruses.Tingting CuiSebastiaan TheunsLowiese M. B. DesmaretsJiexiong XieGaëtan M. A. De GryseBo YangWim Van den BroeckHans J. NauwynckNature PortfolioarticlePorcine Epidemic Diarrhea Virus (PEDV)Transmissible Gastroenteritis Virus (TGEV)PEDV CV777Rotavirus StrainsMilan StrainMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-17 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Porcine Epidemic Diarrhea Virus (PEDV) Transmissible Gastroenteritis Virus (TGEV) PEDV CV777 Rotavirus Strains Milan Strain Medicine R Science Q
spellingShingle	Porcine Epidemic Diarrhea Virus (PEDV) Transmissible Gastroenteritis Virus (TGEV) PEDV CV777 Rotavirus Strains Milan Strain Medicine R Science Q Tingting Cui Sebastiaan Theuns Lowiese M. B. Desmarets Jiexiong Xie Gaëtan M. A. De Gryse Bo Yang Wim Van den Broeck Hans J. Nauwynck Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses
description	Abstract A stable culture of primary porcine enterocytes is necessary to study porcine enteric virus replication characteristics. Because the direct cultivation of primary porcine enterocytes is difficult, alternatives have to be considered. As subepithelial myofibroblasts secrete extracellular matrix and growth factors contributing to the attachment, proliferation and differentiation of epithelial cells, co-cultures of primary porcine enterocytes (ileocytes and colonocytes) with myofibroblasts were developed and evaluated for their susceptibility to enteric viruses. First, it was demonstrated that the co-cultured ileocytes and colonocytes were susceptible to an archival rotavirus strain RVA/pig-tc/BEL/RV277/1977/G1P[7] and different other rotavirus genotypes (fecal samples containing G5P[7], G5P[13], G9P[23], G4P[6]). Next, the TGEV Purdue strain infected both ileocytes and colonocytes whereas the Miller strain only infected ileocytes. Last, the PEDV CV777 Vero adapted and non-adapted (fecal suspension) strains could infect co-cultured ileocytes but not colonocytes. The infectivity of the CV777 Vero adapted strain was higher when the cells were cultured without fetal bovine serum and the CV777 fecal suspension only infected the ileocytes cultured without fetal bovine serum. In conclusion, a novel co-culture of porcine enterocytes with myofibroblasts was established, which can be used for the investigation of the replication of enteric viruses.
format	article
author	Tingting Cui Sebastiaan Theuns Lowiese M. B. Desmarets Jiexiong Xie Gaëtan M. A. De Gryse Bo Yang Wim Van den Broeck Hans J. Nauwynck
author_facet	Tingting Cui Sebastiaan Theuns Lowiese M. B. Desmarets Jiexiong Xie Gaëtan M. A. De Gryse Bo Yang Wim Van den Broeck Hans J. Nauwynck
author_sort	Tingting Cui
title	Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses
title_short	Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses
title_full	Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses
title_fullStr	Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses
title_full_unstemmed	Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses
title_sort	establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/dd10812196a04031b5d536d14c9aa44f
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Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses

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