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Generation of equine enteroids and enteroid-derived 2D monolayers that are responsive to microbial mimics

Hellman, Stina (2021). Generation of equine enteroids and enteroid-derived 2D monolayers that are responsive to microbial mimics. Veterinary Research. 52 , 108
[Research article]

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Abstract

Enteroid cultures are three-dimensional in vitro models that reflect the cellular composition and architecture of the small intestine. One limitation with the enteroid conformation is the enclosed lumen, making it difficult to expose the apical surface of the epithelium to experimental treatments. The present study was therefore conducted to generate cultures of equine enteroids and to develop methods for culture of enteroid-derived cells on a two-dimensional plane, enabling easy access to the apical surface of the epithelium. Equine enteroids were established from small intestinal crypts within 7-9 days of culture. Transcriptional analysis of cell type markers confirmed the presence of enterocytes, stem-, Paneth-, proliferative-, enteroendocrine-, goblet- and tuft cells. This cellular composition was maintained over multiple passages, showing that the enteroids can be kept for prolonged periods. The transfer from 3D enteroids to 2D monolayers slightly modified the relative expression levels of the cell type markers, indicating a decrease of goblet- and Paneth cells in the monolayers. Stimulation with the TLR2, 3 and 4 agonists Pam3CSK4, Poly I:C and LPS, respectively, induced the pro-inflammatory cytokines TNF-alpha and IL-8, while the TLR5 agonist FliC only induced TNF-alpha. In addition, an up-regulation of TGF-beta, IL-33 and IFN-beta was recorded after exposure to lipofected Poly I:C that also affected the monolayer integrity. Thus, the equine enteroid-derived 2D monolayers described in the present study show both genetic and functional similarities with the equine intestine making it an interesting in vitro model for studies demanding access to the apical surface, e.g. in studies of host-microbe interactions.

Authors/Creators:Hellman, Stina
Title:Generation of equine enteroids and enteroid-derived 2D monolayers that are responsive to microbial mimics
Series Name/Journal:Veterinary Research
Year of publishing :2021
Volume:52
Article number:108
Number of Pages:11
Publisher:BMC
ISSN:0928-4249
Language:English
Publication Type:Research article
Article category:Scientific peer reviewed
Version:Published version
Copyright:Creative Commons: Attribution 4.0
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Cell Biology
Keywords:Equine, Enteroid, Enteroid-derived, Epithelium, 2D monolayer, Intestine, In vitro, TLR
URN:NBN:urn:nbn:se:slu:epsilon-p-113439
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-113439
Additional ID:
Type of IDID
DOI10.1186/s13567-021-00976-0
Web of Science (WoS)000686675700001
ID Code:25269
Faculty:VH - Faculty of Veterinary Medicine and Animal Science
Department:(VH) > Department of Biomedical Science and Veterinary Public Health
Deposited By: SLUpub Connector
Deposited On:07 Sep 2021 15:25
Metadata Last Modified:07 Sep 2021 15:31

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