Establishment of fetal bovine intestinal epithelial cell cultures susceptible to bovine rotavirus infection.
Mucosal epithelial cells are infected by a wide variety of pathogens and determining their response to infection is critical for understanding disease pathogenesis. A protocol was developed for culturing primary epithelial cells from fetal bovine intestine and the cultured cells were evaluated for susceptibility to an enteric viral infection. Immunohistochemical staining for cytokeratin confirmed that 60-75% of cultured cells were epithelial cells. Furthermore, following infection with bovine rotavirus (BRV) over 80% of cells in the ileal and jejunal cultures contained viral protein at 16 h post-infection. The intestinal epithelial cell cultures also contained fibroblasts so a jejunal fibroblast culture was established and infected with BRV. Viral protein was detected in jejunal fibroblasts but viral-induced cytopathology was delayed in fibroblast cultures when compared to epithelial cell cultures. This study describes an effective protocol for culturing bovine epithelial cells from fetal intestine and confirmed that the epithelial cells were susceptible to BRV infection. Ileal and jejunal cultures displayed limited growth following continuous passage but early passage epithelial cells provide competent target cells for studying host cell responses to an enteric viral pathogen.
Kaushik
Radhey S
RS
Vaccine & Infectious Disease Organization, University of Saskatchewan, Saskatoon, Saskatchewan, Canada S7N 5E3.
Begg
Ashley A
AA
Wilson
Heather L
HL
Aich
Palok
P
Abrahamsen
Mitchell S
MS
Potter
Andrew
A
Babiuk
Lorne A
LA
Griebel
Philip
P
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000821:virology*; D005347:Fibroblasts / Q000821:virology; D007413:Intestinal Mucosa / Q000166:cytology; D012401:Rotavirus / Q000254:growth & development*; D014776:Virus Cultivation / Q000379:methods*