Establishment of fetal bovine intestinal epithelial cell cultures susceptible to bovine rotavirus infection

Journal of Virological Methods
Radhey S KaushikPhilip Griebel

Abstract

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 studyin...Continue Reading

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Citations

Oct 16, 2009·Histochemistry and Cell Biology·Kohtaro MiyazawaHisashi Aso
Aug 29, 2013·In Vitro Cellular & Developmental Biology. Animal·Ling GongWei Li
Jun 1, 2010·Probiotics and Antimicrobial Proteins·Alison ThompsonPalok Aich
Apr 21, 2010·Journal of Virological Methods·Carlos A GuerreroOrlando Acosta
Jul 5, 2012·Journal of Peptide Science : an Official Publication of the European Peptide Society·Besnik KrasniqiScott Napper
Jun 29, 2012·The Journal of General Virology·Zhi SunYing Fang
Mar 20, 2014·Journal of Virological Methods·Jing WangYanming Zhang
Feb 14, 2015·Physiological Reports·Pu MaoYimin Li
Jun 12, 2019·In Vitro Cellular & Developmental Biology. Animal·Tirth UpretyRadhey S Kaushik
Dec 2, 2020·Comparative Immunology, Microbiology and Infectious Diseases·Pratik KatwalRadhey S Kaushik
Mar 14, 2021·Veterinary Immunology and Immunopathology·Brian M KobylkevichMark A Messerli

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Methods Mentioned

BETA
PCR
flow cytometry
dissection
FCS

Software Mentioned

CellQuest

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