Development of in vitro enteroids derived from bovine small intestinal crypts

Hamilton, C. A. et al. (2018) Development of in vitro enteroids derived from bovine small intestinal crypts. Veterinary Research, 49, 54. (doi: 10.1186/s13567-018-0547-5) (PMID:29970174) (PMCID:PMC6029049)

Abstract

Cattle are an economically important domestic animal species. In vitro 2D cultures of intestinal epithelial cells or epithelial cell lines have been widely used to study cell function and host-pathogen interactions in the bovine intestine. However, these cultures lack the cellular diversity encountered in the intestinal epithelium, and the physiological relevance of monocultures of transformed cell lines is uncertain. Little is also known of the factors that influence cell differentiation and homeostasis in the bovine intestinal epithelium, and few cell-specific markers that can distinguish the different intestinal epithelial cell lineages have been reported. Here we describe a simple and reliable procedure to establish in vitro 3D enteroid, or "mini gut", cultures from bovine small intestinal (ileal) crypts. These enteroids contained a continuous central lumen lined with a single layer of polarized enterocytes, bound by tight junctions with abundant microvilli on their apical surfaces. Histological and transcriptional analyses suggested that the enteroids comprised a mixed population of intestinal epithelial cell lineages including intestinal stem cells, enterocytes, Paneth cells, goblet cells and enteroendocrine cells. We show that bovine enteroids can be successfully maintained long-term through multiple serial passages without observable changes to their growth characteristics, morphology or transcriptome. Furthermore, the bovine enteroids can be cryopreserved and viable cultures recovered from frozen stocks. Our data suggest that these 3D bovine enteroid cultures represent a novel, physiologically-relevant and tractable in vitro system in which epithelial cell differentiation and function, and host-pathogen interactions in the bovine small intestine can be studied.

• Item Type: Articles
• Additional Information: This study was funded by Biotechnology and Biological Sciences Research Council (GB; grant references BB/M012751/1, BB/ M012808/1, BBS/E/D/20231762 and BBS/E/D/20002174); Royal Society (grant reference UF140610).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Sehgal, Dr Anuj
• Authors: Hamilton, C. A., Young, R., Jayaraman, S., Sehgal, A., Paxton, E., Thomson, S., Katzer, F., Hope, J., Innes, E., Morrison, L. J., and Mabbott, N. A.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Veterinary Research
• Publisher: BioMed Central
• ISSN: 0928-4249
• ISSN (Online): 1297-9716
• Copyright Holders: Copyright © 2018 The Authors
• First Published: First published in Veterinary Research 49: 54
• Publisher Policy: Reproduced under a Creative Commons License