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

Tingting Cui (UGent) , Sebastiaan Theuns (UGent) , Lowiese Desmarets (UGent) , Jiexiong Xie (UGent) , Gaëtan De Gryse (UGent) , Bo Yang (UGent) , Wim Van Den Broeck (UGent) and Hans Nauwynck (UGent)
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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.
Keywords
TRANSMISSIBLE GASTROENTERITIS VIRUS, GROUP-A ROTAVIRUSES, AMINOPEPTIDASE-N, INTESTINAL MYOFIBROBLASTS, EPIDEMIC DIARRHEA, EPITHELIAL-CELLS, HUMAN ENTEROIDS, MAJOR RECEPTOR, SPIKE PROTEIN, TISSUE-REPAIR

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Chicago
Cui, Tingting, Sebastiaan Theuns, Lowiese Desmarets, Jiexiong Xie, Gaëtan De Gryse, Bo Yang, Wim Van Den Broeck, and Hans Nauwynck. 2018. “Establishment of Porcine Enterocyte/myofibroblast Co-cultures for the Growth of Porcine Rota- and Coronaviruses.” Scientific Reports 8.
APA
Cui, T., Theuns, S., Desmarets, L., Xie, J., De Gryse, G., Yang, B., Van Den Broeck, W., et al. (2018). Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses. SCIENTIFIC REPORTS, 8.
Vancouver
1.
Cui T, Theuns S, Desmarets L, Xie J, De Gryse G, Yang B, et al. Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses. SCIENTIFIC REPORTS. 2018;8.
MLA
Cui, Tingting et al. “Establishment of Porcine Enterocyte/myofibroblast Co-cultures for the Growth of Porcine Rota- and Coronaviruses.” SCIENTIFIC REPORTS 8 (2018): n. pag. Print.
@article{8598770,
  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.},
  articleno    = {15195},
  author       = {Cui, Tingting and Theuns, Sebastiaan and Desmarets, Lowiese and Xie, Jiexiong and De Gryse, Ga{\"e}tan and Yang, Bo and Van Den Broeck, Wim and Nauwynck, Hans},
  issn         = {2045-2322},
  journal      = {SCIENTIFIC REPORTS},
  language     = {eng},
  pages        = {17},
  title        = {Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses},
  url          = {http://dx.doi.org/10.1038/s41598-018-33305-1},
  volume       = {8},
  year         = {2018},
}

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