In‐depth analysis of the relationship between bovine intestinal organoids and enteroids based on morphology and transcriptome

Abstract

Intestinal organoids and enteroids as excellent models are miniaturized and simplified for studying intestinal physiological and pathological functions, drug screening, and regenerative medicine. Recently, the application demands for organoids and enteroids in organ development and nutrition metabolism, immune and cancer research increased. But there are few comparative studies on both of them, especially in immunity and metabolism, which is also conducive to further clarifying the role of crypt stem cells and stromal cells. In our study, “natural” organoids were obtained by tissue culture from fetal bovine jejunum and enteroids were successfully isolated and cultured from organoids without supplementing exogenous factors and Matrigel. These mini‐guts displayed similar features to the intestine through immunohistochemistry and transmission electron microscopy. Organoid and enteroid were systematically compared based on the transcriptome. And some of the results were verified by qRT‐PCR. Our results showed KDGs (Key driver genes) (e.g., SLC13A1, HOXA7, HOXA6, HOXA5, and HOXD4) of organoids enriched in signaling pathways related to organ development and morphology and metabolism. KDGs (e.g., IL‐6, PTGS2, CDH1, JUN, and EGFR) of enteroid were involved in cancer, MAPK, and immune‐related signaling pathways. To the Wnt signaling pathway, highly expressed genes in organoids, including RSPO2, NOTUM, WNT6, and RSPO3, supported the homeostasis of crypt stem cells. Enteroids highly expressed CTNNB1 and WNTs. In addition, we found that organoids and enteroids carried out different functions in immunity and metabolism due to different cell compositions. Therefore, it suggested organoid is more compatible and comprehensive, and enteroid is qualified for the research of immunity and cancer.

Adult Bovine-Derived Small and Large Intestinal Organoids: In Vitro Development and Maintenance

“…As a limitation, the present study did not evaluate the direct gene expression of the donor tissue; thus, it is not possible to compare and discuss similarities or diferences in genetic properties between the organoids and their originating tissues. However, previous studies reported an overlapping gene expression pattern between bovine intestinal organoids and the in vivo tissue [15,16,44]. Tis characteristic has also been reported for other types of organoids such as the human and murine liver [45,46].…”

Bovine Enteroids as an In Vitro Model for Infection with Bovine Coronavirus

“…In the present study, several formulations ( Table S1 ) for the manipulation of growth factors and differentiation of the enteroids were tested, but our results did not indicate that modifying the concentration of growth factors mentioned above was necessary for differentiation. Embedding the enteroids in Matrigel could also have contributed to differentiation, as described by [ 51 , 52 ]. To lower any Matrigel-driven differentiation, we used Matrigel with reduced growth factors [ 18 ].…”

Modeling Paratuberculosis in Laboratory Animals, Cells, or Tissues: A Focus on Their Applications for Pathogenesis, Diagnosis, Vaccines, and Therapy Studies

“…The protocols for culturing ruminant organoids from different digestive segments have been set out in [112][113][114][115][116][117][118][119][120]. Since the first report in 2009, intestinal organoids have evolved as a potential alternative to in vivo models for various experimental purposes, such as imaging, molecular analysis and gene editing, but also as reductionist approaches with which to study the interaction of epithelial cells with the other relevant actors in a physiological context, such as the immune cells [121,122] or microbiota [123,124], or even to evaluate the early host-pathogen interaction in the context of many infectious diseases (protozoa [125,126], virus [127], and bacteria [128][129][130][131][132][133]).…”