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Establishment of porcine enterocyte/myofibroblast co-cultures for the growth of porcine rota- and coronaviruses.
- Source :
-
Scientific reports [Sci Rep] 2018 Oct 12; Vol. 8 (1), pp. 15195. Date of Electronic Publication: 2018 Oct 12. - Publication Year :
- 2018
-
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.
- Subjects :
- Animals
Colon pathology
Colon virology
Diarrhea virology
Enterocytes pathology
Epithelial Cells ultrastructure
Epithelial Cells virology
Feces virology
Genotype
Ileum pathology
Ileum virology
Kinetics
Myofibroblasts pathology
Rotavirus genetics
Virus Replication
Coculture Techniques methods
Coronavirus growth & development
Enterocytes virology
Myofibroblasts virology
Rotavirus growth & development
Swine virology
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 8
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 30315177
- Full Text :
- https://doi.org/10.1038/s41598-018-33305-1