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

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 studying host cell responses to an enteric viral pathogen.

Characterization of newly established bovine intestinal epithelial cell line

“…Additionally, Kaushik et al have succeeded in cultivating primary bovine intestinal epithelial cells from fetal intestine (Kaushik et al 2008). We therefore considered that fetal material might be useful for the establishment of an intestinal epithelial cell line.…”

“…Some groups have reported the establishment of bovine primary intestinal epithelial cell cultures (Follmann et al 2000;Kaushik et al 2008) and cloned bovine jejunocyte and colonocyte cell lines have been established (Loret et al 2009). Cloned cell lines have the advantage over primary cultures in that they retain reasonably constant characteristics for numerous passages.…”

Probiotic-Induced Priming of Innate Immunity to Protect Against Rotaviral Infection

“…In-house BIECs were developed at VIDO as described by Kaushik et al [11]. Adaptation of the primary cell line yielded a more sustainable culture, known as EVM07-BIEC.…”

“…Over 25 years have passed since it was first proposed that a stable intestinal epithelial line should be developed and the effects of passive immunity in preventing a rotavirus infection should be researched [16]. We have developed this in-house stable line of bovine intestinal epithelial cells (BIEC) that is susceptible to rotavirus infection [2,11]. Bovine rotavirus (BRV) causes an enteric infection in newborn calves up to approximately 3 weeks of age [16].…”

Establishment and Characterization of an SV40 Large T Antigen-Transduced Porcine Colonic Epithelial Cell Line

“…Well-established protocols are available for the isolation and cultivation of intestinal epithelial cells derived from different species that maintain the characteristics of the original cells [Golaz et al, 2007;Kaushik et al, 2008;Petto et al, 2011;Ren et al, 2011]. Nevertheless, the use of primary cultures is limited by small proliferative life spans that precede permanent growth arrest, termed "in vitro cellular senescence" [Hayflick, 1965[Hayflick, , 1985Kuilman et al, 2010;Rodier and Campisi, 2011].…”