FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Veterinary research2024; 55(1); 30; doi: 10.1186/s13567-024-01283-0

A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium.

Abstract: Epithelial damage due to gastrointestinal disorders frequently causes severe disease in horses. To study the underlying pathophysiological processes, we aimed to establish equine jejunum and colon enteroids (eqJE, eqCE) mimicking the in vivo epithelium. Therefore, enteroids were cultivated in four different media for differentiation and subsequently characterized histomorphologically, on mRNA and on protein level in comparison to the native epithelium of the same donor horses to identify ideal culture conditions for an in vitro model system. With increasing enterocyte differentiation, the enteroids showed a reduced growth rate as well as a predominantly spherical morphology and less budding compared to enteroids in proliferation medium. Combined or individual withdrawal of stem cell niche pathway components resulted in lower mRNA expression levels of stem cell markers and concomitant differentiation of enterocytes, goblet cells and enteroendocrine cells. For eqCE, withdrawal of Wnt alone was sufficient for the generation of differentiated enterocytes with a close resemblance to the in vivo epithelium. Combined removal of Wnt, R-spondin and Noggin and the addition of DAPT stimulated differentiation of eqJE at a similar level as the in vivo epithelium, particularly with regard to enterocytes. In summary, we successfully defined a medium composition that promotes the formation of eqJE and eqCE consisting of multiple cell types and resembling the in vivo epithelium. Our findings emphasize the importance of adapting culture conditions to the respective species and the intestinal segment. This in vitro model will be used to investigate the pathological mechanisms underlying equine gastrointestinal disorders in future studies.
© 2024. The Author(s).
Get Full Text
Publication Date: 2024-03-16 PubMed ID: 38493107PubMed Central: 4777998DOI: 10.1186/s13567-024-01283-0Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The researchers successfully developed a lab-grown model of horse intestine tissue, known as enteroids, to study how gastrointestinal diseases affect horses. They also highlighted the need for species- and region-specific culture conditions when constructing these models.

Objective of the Study

  • The researchers aimed to create a model of horse intestinal tissue (specifically jejunum and colon sections, labeled eqJE and eqCE respectively) in a lab setting, to understand how gastrointestinal disorders damage horse tissues.

Methodology

  • Enteroids were cultivated in four different types of media to ensure differentiation (the process by which cells become more specialized).
  • These lab-grown tissues were then compared to actual tissue samples from the same horses, to check how closely the former emulated the latter. This comparison was done on a histomorphological level (study of the form and structure of the tissues), mRNA level (gene expression), and protein level.

Key Findings

  • As the enteroids matured and further specialized (captured by increasing enterocyte differentiation), the tissues slowed their growth and formed mostly round shapes with fewer outgrowths than those in proliferation medium (conditions that promote growth).
  • When stem cell niche pathway components were reduced or removed individually or collectively, the enteroids displayed lowered expression levels of stem cell markers. These removals also induced further differentiation in the enterocytes, goblet cells, and enteroendocrine cells.
  • The removal of the Wnt protein alone resulted in eqCE generating differentiated enterocytes similar to the natural tissues. Meanwhile, removing Wnt, R-spondin, and Noggin, and adding DAPT resulted in eqJE differentiating at similar levels.

Implications

  • The researchers successfully created a culture medium that encouraged the differentiation of eqJE and eqCE into multiple cell types. These tissues closely resembled their in vivo counterpart.
  • The study underlined the importance of tailoring culture conditions to the specific species and intestinal segment being grown.
  • This newly-developed in vitro model will now be used to delve deeper into the pathological mechanisms at play in equine gastrointestinal disorders.

Cite This Article

APA
Windhaber C, Heckl A, Csukovich G, Pratscher B, Burgener IA, Biermann N, Dengler F. (2024). A matter of differentiation: equine enteroids as a model for the in vivo intestinal epithelium. Vet Res, 55(1), 30. https://doi.org/10.1186/s13567-024-01283-0

Publication

Veterinary research
ISSN: 1297-9716
NlmUniqueID: 9309551
Country: England
Language: English
Volume: 55
Issue: 1
Pages: 30

Researcher Affiliations

Windhaber, Christina
  • Institute of Physiology, Pathophysiology and Biophysics, University of Veterinary Medicine, Vienna, Austria.
Heckl, Anna
  • Institute of Physiology, Pathophysiology and Biophysics, University of Veterinary Medicine, Vienna, Austria.
Csukovich, Georg
  • Division of Small Animal Internal Medicine, University of Veterinary Medicine, Vienna, Austria.
Pratscher, Barbara
  • Division of Small Animal Internal Medicine, University of Veterinary Medicine, Vienna, Austria.
Burgener, Iwan Anton
  • Division of Small Animal Internal Medicine, University of Veterinary Medicine, Vienna, Austria.
Biermann, Nora
  • Clinical Unit of Equine Surgery, University of Veterinary Medicine, Vienna, Austria.
Dengler, Franziska
  • Institute of Physiology, Pathophysiology and Biophysics, University of Veterinary Medicine, Vienna, Austria. franziska.dengler@vetmeduni.ac.at.

MeSH Terms

  • Animals
  • Horses
  • Intestinal Mucosa
  • Intestines
  • Cell Differentiation
  • Gastrointestinal Diseases / veterinary
  • RNA, Messenger
  • Horse Diseases

References

This article includes 74 references
  1. Tinker MK, White NA, Lessard P, Thatcher CD, Pelzer KD, Davis B, Carmel DK. Prospective study of equine colic incidence and mortality.. Equine Vet J 29:448–453.
    pubmed: 9413717doi: 10.1111/j.2042-3306.1997.tb03157.xgoogle scholar: lookup
  2. Ireland JL, Clegg PD, McGowan CM, Platt L, Pinchbeck GL. Factors associated with mortality of geriatric horses in the United Kingdom.. Prev Vet Med 101:204–218.
    pubmed: 21733586doi: 10.1016/j.prevetmed.2011.06.002google scholar: lookup
  3. van der Linden MA, Laffont CM, van Sloet Oldruitenborgh-Oosterbaan MM. Prognosis in equine medical and surgical colic.. J Vet Intern Med 17:343–348.
    pubmed: 12774977doi: 10.1111/j.1939-1676.2003.tb02459.xgoogle scholar: lookup
  4. Grootjans J, Lenaerts K, Buurman WA, Dejong CHC, Derikx JPM. Life and death at the mucosal-luminal interface: new perspectives on human intestinal ischemia-reperfusion.. World J Gastroenterol 22:2760–2770.
    pubmed: 26973414pmc: 4777998doi: 10.3748/wjg.v22.i9.2760google scholar: lookup
  5. Lechuga S, Ivanov AI. Disruption of the epithelial barrier during intestinal inflammation: quest for new molecules and mechanisms.. Biochim Biophys Acta Mol Cell Res 1864:1183–1194.
    pubmed: 28322932doi: 10.1016/j.bbamcr.2017.03.007google scholar: lookup
  6. Blikslager A, Gonzalez L. Equine intestinal mucosal pathobiology.. Annu Rev Anim Biosci 6:157–175.
    pubmed: 29144770doi: 10.1146/annurev-animal-030117-014748google scholar: lookup
  7. Martini E, Krug SM, Siegmund B, Neurath MF, Becker C. Mend your fences: the epithelial barrier and its relationship with mucosal immunity in inflammatory bowel disease.. Cell Mol Gastroenterol Hepatol 4:33–46.
    pubmed: 28560287pmc: 5439240doi: 10.1016/j.jcmgh.2017.03.007google scholar: lookup
  8. Umar S. Intestinal stem cells.. Curr Gastroenterol Rep 12:340–348.
    pubmed: 20683682pmc: 2965634doi: 10.1007/s11894-010-0130-3google scholar: lookup
  9. Sambuy Y, de Angelis I, Ranaldi G, Scarino ML, Stammati A, Zucco F. The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics.. Cell Biol Toxicol 21:1–26.
    pubmed: 15868485doi: 10.1007/s10565-005-0085-6google scholar: lookup
  10. Noben M, Vanhove W, Arnauts K, Santo Ramalho A, van Assche G, Vermeire S, Verfaillie C, Ferrante M. Human intestinal epithelium in a dish: current models for research into gastrointestinal pathophysiology.. United European Gastroenterol J 5:1073–1081.
    pubmed: 29238585pmc: 5721984doi: 10.1177/2050640617722903google scholar: lookup
  11. Quaroni A, Wands J, Trelstad RL, Isselbacher KJ. Epithelioid cell cultures from rat small intestine. Characterization by morphologic and immunologic criteria.. J Cell Biol 80:248–265.
    pubmed: 88453doi: 10.1083/jcb.80.2.248google scholar: lookup
  12. Bartsch I, Zschaler I, Haseloff M, Steinberg P. Establishment of a long-term culture system for rat colon epithelial cells.. In Vitro Cell Dev Biol Anim 40:278–284.
    pubmed: 15723563doi: 10.1290/0404035.1google scholar: lookup
  13. Weng X-H, Beyenbach KW, Quaroni A. Cultured monolayers of the dog jejunum with the structural and functional properties resembling the normal epithelium.. Am J Physiol Gastrointest Liver Physiol 288:G705–G717.
    pubmed: 15550553doi: 10.1152/ajpgi.00518.2003google scholar: lookup
  14. van der Flier LG, Clevers H. Stem cells, self-renewal, and differentiation in the intestinal epithelium.. Annu Rev Physiol 71:241–260.
    pubmed: 18808327doi: 10.1146/annurev.physiol.010908.163145google scholar: lookup
  15. Zachos NC, Kovbasnjuk O, Foulke-Abel J, In J, Blutt SE, de Jonge HR, Estes MK, Donowitz M. Human enteroids/colonoids and intestinal organoids functionally recapitulate normal intestinal physiology and pathophysiology.. J Biol Chem 291:3759–3766.
    pubmed: 26677228doi: 10.1074/jbc.R114.635995google scholar: lookup
  16. Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE, van Es JH, Abo A, Kujala P, Peters PJ, Clevers H. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche.. Nature 459:262–265.
    pubmed: 19329995doi: 10.1038/nature07935google scholar: lookup
  17. Jung P, Sato T, Merlos-Suárez A, Barriga FM, Iglesias M, Rossell D, Auer H, Gallardo M, Blasco MA, Sancho E, Clevers H, Batlle E. Isolation and in vitro expansion of human colonic stem cells.. Nat Med 17:1225–1227.
    pubmed: 21892181doi: 10.1038/nm.2470google scholar: lookup
  18. Pinto D, Clevers H. Wnt control of stem cells and differentiation in the intestinal epithelium.. Exp Cell Res 306:357–363.
    pubmed: 15925592doi: 10.1016/j.yexcr.2005.02.022google scholar: lookup
  19. Sato T, Clevers H. Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications.. Science 340:1190–1194.
    pubmed: 23744940doi: 10.1126/science.1234852google scholar: lookup
  20. Tsai YH, Wu A, Wu JH, Capeling MM, Holloway EM, Huang S, Czerwinkski M, Glass I, Higgins PDR, Spence JR. Acquisition of NOTCH dependence is a hallmark of human intestinal stem cell maturation.. Stem Cell Reports 17:1138–1153.
    pubmed: 35395175pmc: 9133587doi: 10.1016/j.stemcr.2022.03.007google scholar: lookup
  21. van Es JH, van Gijn ME, Riccio O, van den Born M, Vooijs M, Begthel H, Cozijnsen M, Robine S, Winton DJ, Radtke F, Clevers H. Notch/gamma-secretase inhibition turns proliferative cells in intestinal crypts and adenomas into goblet cells.. Nature 435:959–963.
    pubmed: 15959515doi: 10.1038/nature03659google scholar: lookup
  22. Li Y, Liu Y, Liu B, Wang J, Wei S, Qi Z, Wang S, Fu W, Chen YG. A growth factor-free culture system underscores the coordination between Wnt and BMP signaling in Lgr5 intestinal stem cell maintenance.. Cell Discov 4:49.
    pubmed: 30181900pmc: 6120946doi: 10.1038/s41421-018-0051-0google scholar: lookup
  23. Sato T, Stange DE, Ferrante M, Vries RG, Van Es JH, Van den Brink S, Van Houdt WJ, Pronk A, Van Gorp J, Siersema PD, Clevers H. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium.. Gastroenterology 141:1762–1772.
    pubmed: 21889923doi: 10.1053/j.gastro.2011.07.050google scholar: lookup
  24. Töpfer E, Pasotti A, Telopoulou A, Italiani P, Boraschi D, Ewart M-A, Wilde C. Bovine colon organoids: From 3D bioprinting to cryopreserved multi-well screening platforms.. Toxicol In Vitro 61:104606.
    pubmed: 31344400doi: 10.1016/j.tiv.2019.104606google scholar: lookup
  25. VanDussen KL, Marinshaw JM, Shaikh N, Miyoshi H, Moon C, Tarr PI, Ciorba MA, Stappenbeck TS. Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays.. Gut 64:911–920.
    pubmed: 25007816doi: 10.1136/gutjnl-2013-306651google scholar: lookup
  26. Wilson SS, Mayo M, Melim T, Knight H, Patnaude L, Wu X, Phillips L, Westmoreland S, Dunstan R, Fiebiger E, Terrillon S. Optimized culture conditions for improved growth and functional differentiation of mouse and human colon organoids.. Front Immunol 11:547102.
    pubmed: 33643277doi: 10.3389/fimmu.2020.547102google scholar: lookup
  27. Mussard E, Pouzet C, Helies V, Pascal G, Fourre S, Cherbuy C, Rubio A, Vergnolle N, Combes S, Beaumont M. Culture of rabbit caecum organoids by reconstituting the intestinal stem cell niche in vitro with pharmacological inhibitors or L-WRN conditioned medium.. Stem Cell Res 48:101980.
    pubmed: 32920507doi: 10.1016/j.scr.2020.101980google scholar: lookup
  28. Kip AM, Soons Z, Mohren R, Duivenvoorden AAM, Röth AAJ, Cillero-Pastor B, Neumann UP, Dejong CHC, Heeren RMA, Olde Damink SWM, Lenaerts K. Proteomics analysis of human intestinal organoids during hypoxia and reoxygenation as a model to study ischemia-reperfusion injury.. Cell Death Dis 12:95.
    pubmed: 33462215pmc: 7813872doi: 10.1038/s41419-020-03379-9google scholar: lookup
  29. Seeger B. Farm animal-derived models of the intestinal epithelium: recent advances and future applications of intestinal organoids.. Altern Lab Anim 48:215–233.
    pubmed: 33337913doi: 10.1177/0261192920974026google scholar: lookup
  30. Rahmani S, Breyner NM, Su H-M, Verdu EF, Didar TF. Intestinal organoids: a new paradigm for engineering intestinal epithelium in vitro.. Biomaterials 194:195–214.
    pubmed: 30612006doi: 10.1016/j.biomaterials.2018.12.006google scholar: lookup
  31. Pleguezuelos-Manzano C, Puschhof J, van den Brink S, Geurts V, Beumer J, Clevers H. Establishment and culture of human intestinal organoids derived from adult stem cells.. Curr Protoc Immunol 130:e106.
    pubmed: 32940424pmc: 9285512doi: 10.1002/cpim.106google scholar: lookup
  32. Wang Y, Kim R, Gunasekara DB, Reed MI, DiSalvo M, Nguyen DL, Bultman SJ, Sims CE, Magness ST, Allbritton NL. Formation of human colonic crypt array by application of chemical gradients across a shaped epithelial monolayer.. Cell Mol Gastroenterol Hepatol 5:113–130.
    pubmed: 29693040doi: 10.1016/j.jcmgh.2017.10.007google scholar: lookup
  33. Miyoshi H, Stappenbeck TS. In vitro expansion and genetic modification of gastrointestinal stem cells in spheroid culture.. Nat Protoc 8:2471–2482.
    pubmed: 24232249pmc: 3969856doi: 10.1038/nprot.2013.153google scholar: lookup
  34. Kramer N, Pratscher B, Meneses AMC, Tschulenk W, Walter I, Swoboda A, Kruitwagen HS, Schneeberger K, Penning LC, Spee B, Kieslinger M, Brandt S, Burgener IA. Generation of differentiating and long-living intestinal organoids reflecting the cellular diversity of canine intestine.. Cells 9:822.
    pubmed: 32231153pmc: 7226743doi: 10.3390/cells9040822google scholar: lookup
  35. Barnett AM, Mullaney JA, Hendriks C, Le Borgne L, McNabb WC, Roy NC. Porcine colonoids and enteroids keep the memory of their origin during regeneration.. Am J Physiol Cell Physiol 320:C794–C805.
    pubmed: 33760661doi: 10.1152/ajpcell.00420.2020google scholar: lookup
  36. Gonzalez LM, Williamson I, Piedrahita JA, Blikslager AT, Magness ST. Cell lineage identification and stem cell culture in a porcine model for the study of intestinal epithelial regeneration.. PLoS ONE 8:e66465.
    pubmed: 23840480pmc: 3696067doi: 10.1371/journal.pone.0066465google scholar: lookup
  37. Hamilton CA, Young R, Jayaraman S, Sehgal A, Paxton E, Thomson S, Katzer F, Hope J, Innes E, Morrison LJ, Mabbott NA. Development of in vitro enteroids derived from bovine small intestinal crypts.. Vet Res 49:54.
    pubmed: 29970174pmc: 6029049doi: 10.1186/s13567-018-0547-5google scholar: lookup
  38. Khalil HA, Lei NY, Brinkley G, Scott A, Wang J, Kar UK, Jabaji ZB, Lewis M, Martín MG, Dunn JC, Stelzner MG. A novel culture system for adult porcine intestinal crypts.. Cell Tissue Res 365:123–134.
    pubmed: 26928041pmc: 4919165doi: 10.1007/s00441-016-2367-0google scholar: lookup
  39. Nash TJ, Morris KM, Mabbott NA, Vervelde L. Inside-out chicken enteroids with leukocyte component as a model to study host-pathogen interactions.. Commun Biol 4:377.
    pubmed: 33742093pmc: 7979936doi: 10.1038/s42003-021-01901-zgoogle scholar: lookup
  40. Blake R, Jensen K, Mabbott N, Hope J, Stevens J. The development of 3D bovine intestinal organoid derived models to investigate Mycobacterium Avium ssp Paratuberculosis pathogenesis.. Front Vet Sci 9:921160.
    pubmed: 35859809pmc: 9290757doi: 10.3389/fvets.2022.921160google scholar: lookup
  41. Sutton KM, Orr B, Hope J, Jensen SR, Vervelde L. Establishment of bovine 3D enteroid-derived 2D monolayers.. Vet Res 53:15.
    pubmed: 35236416pmc: 8889782doi: 10.1186/s13567-022-01033-0google scholar: lookup
  42. Hoffmann P, Schnepel N, Langeheine M, Künnemann K, Grassl GA, Brehm R, Seeger B, Mazzuoli-Weber G, Breves G. Intestinal organoid-based 2D monolayers mimic physiological and pathophysiological properties of the pig intestine.. PLoS ONE 16:e0256143.
    pubmed: 34424915pmc: 8382199doi: 10.1371/journal.pone.0256143google scholar: lookup
  43. van der Hee B, Madsen O, Vervoort J, Smidt H, Wells JM. Congruence of transcription programs in adult stem cell-derived jejunum organoids and original tissue during long-term culture.. Front Cell Dev Biol 8:375.
    pubmed: 32714922pmc: 7343960doi: 10.3389/fcell.2020.00375google scholar: lookup
  44. Stieler Stewart A, Freund JM, Gonzalez LM. Advanced three-dimensional culture of equine intestinal epithelial stem cells.. Equine Vet J 50:241–248.
    pubmed: 28792626doi: 10.1111/evj.12734google scholar: lookup
  45. Hellman S. Generation of equine enteroids and enteroid-derived 2D monolayers that are responsive to microbial mimics.. Vet Res 52:108.
    pubmed: 34391473pmc: 8364015doi: 10.1186/s13567-021-00976-0google scholar: lookup
  46. Stewart AS, Schaaf CR, Veerasammy B, Freund JM, Gonzalez LM. Culture of equine intestinal epithelial stem cells after delayed tissue storage for future applications.. BMC Vet Res 18:445.
    pubmed: 36564773pmc: 9783463doi: 10.1186/s12917-022-03552-6google scholar: lookup
  47. Hellman S. New dimensions of equine intestinal organoids to model host-microbe interactions in vitro.. Equine Vet J 53:34.
    doi: 10.1111/evj.41_13495google scholar: lookup
  48. O’Rourke K, Ackerman S, Dow L, Lowe S. Isolation, culture, and maintenance of mouse intestinal stem cells.. Bio Protoc 6:e1733.
    pubmed: 27570799
  49. Pastuła A, Middelhoff M, Brandtner A, Tobiasch M, Höhl B, Nuber AH, Demir IE, Neupert S, Kollmann P, Mazzuoli-Weber G, Quante M. Three-dimensional gastrointestinal organoid culture in combination with nerves or fibroblasts: a method to characterize the gastrointestinal stem cell niche.. Stem Cells Int 2016:3710836.
    pubmed: 26697073doi: 10.1155/2016/3710836google scholar: lookup
  50. Nam MO, Hahn S, Jee JH, Hwang TS, Yoon H, Lee DH, Kwon MS, Yoo J. Effects of a small molecule R-spondin-1 substitute RS-246204 on a mouse intestinal organoid culture.. Oncotarget 9:6356–6368.
    pubmed: 29464078doi: 10.18632/oncotarget.23721google scholar: lookup
  51. Basak O, Beumer J, Wiebrands K, Seno H, van Oudenaarden A, Clevers H. Induced quiescence of Lgr5 stem cells in intestinal organoids enables differentiation of hormone-producing enteroendocrine cells.. Cell Stem Cell 20:177-190.e4.
    pubmed: 27939219doi: 10.1016/j.stem.2016.11.001google scholar: lookup
  52. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A. Fiji: an open-source platform for biological-image analysis.. Nat Methods 9:676–682.
    pubmed: 22743772doi: 10.1038/nmeth.2019google scholar: lookup
  53. Dengler F, Sternberg F, Grages M, Kästner SB, Verhaar N. Adaptive mechanisms in no flow vs low flow ischemia in equine Jejunum epithelium: different paths to the same destination.. Front Vet Sci 9:947482.
    pubmed: 36157182pmc: 9493374doi: 10.3389/fvets.2022.947482google scholar: lookup
  54. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments.. Clin Chem 55:611–622.
    pubmed: 19246619doi: 10.1373/clinchem.2008.112797google scholar: lookup
  55. Xie F, Wang J, Zhang B. RefFinder: a web-based tool for comprehensively analyzing and identifying reference genes.. Funct Integr Genomics 23:125.
    pubmed: 37060478doi: 10.1007/s10142-023-01055-7google scholar: lookup
  56. van der Flier LG, Haegebarth A, Stange DE, van de Wetering M, Clevers H. OLFM4 is a robust marker for stem cells in human intestine and marks a subset of colorectal cancer cells.. Gastroenterology 137:15–17.
    pubmed: 19450592doi: 10.1053/j.gastro.2009.05.035google scholar: lookup
  57. Powell RH, Behnke MS. WRN conditioned media is sufficient for in vitro propagation of intestinal organoids from large farm and small companion animals.. Biol Open 6:698–705.
    pubmed: 28347989pmc: 5450310
  58. Ohira S, Yokoi Y, Ayabe T, Nakamura K. Efficient and simple genetic engineering of enteroids using mouse isolated crypts for investigating intestinal functions.. Biochem Biophys Res Commun 637:153–160.
    pubmed: 36402064doi: 10.1016/j.bbrc.2022.11.008google scholar: lookup
  59. Clevers HC, Bevins CL. Paneth cells: maestros of the small intestinal crypts.. Annu Rev Physiol 75:289–311.
    pubmed: 23398152doi: 10.1146/annurev-physiol-030212-183744google scholar: lookup
  60. Holly M, Smith J. Paneth cells during viral infection and pathogenesis.. Viruses 10:225.
    pubmed: 29701691pmc: 5977218doi: 10.3390/v10050225google scholar: lookup
  61. de Lange IH, van Gorp C, Massy KRI, Kessels L, Kloosterboer N, Bjørnshave A, Stampe Ostenfeld M, Damoiseaux JGMC, Derikx JPM, van Gemert WG, Wolfs TGAM. Hypoxia-driven changes in a human intestinal organoid model and the protective effects of hydrolyzed whey.. Nutrients 15:393.
    pubmed: 36678267pmc: 9863820doi: 10.3390/nᔂ0393google scholar: lookup
  62. Pearce SC, Al-Jawadi A, Kishida K, Yu S, Hu M, Fritzky LF, Edelblum KL, Gao N, Ferraris RP. Marked differences in tight junction composition and macromolecular permeability among different intestinal cell types.. BMC Biol 16:19.
    pubmed: 29391007pmc: 5793346doi: 10.1186/s12915-018-0481-zgoogle scholar: lookup
  63. Chandra L, Borcherding DC, Kingsbury D, Atherly T, Ambrosini YM, Bourgois-Mochel A, Yuan W, Kimber M, Qi Y, Wang Q, Wannemuehler M, Ellinwood NM, Snella E, Martin M, Skala M, Meyerholz D, Estes M, Fernandez-Zapico ME, Jergens AE, Mochel JP, Allenspach K. Derivation of adult canine intestinal organoids for translational research in gastroenterology.. BMC Biol 17:33.
    pubmed: 30975131pmc: 6460554doi: 10.1186/s12915-019-0652-6google scholar: lookup
  64. Barker N, van Es JH, Kuipers J, Kujala P, van den Born M, Cozijnsen M, Haegebarth A, Korving J, Begthel H, Peters PJ, Clevers H. Identification of stem cells in small intestine and colon by marker gene Lgr5.. Nature 449:1003–1007.
    pubmed: 17934449doi: 10.1038/nature06196google scholar: lookup
  65. Gonzalez LM, Kinnin LA, Blikslager AT. Characterization of discrete equine intestinal epithelial cell lineages.. Am J Vet Res 76:358–366.
    pubmed: 25815577pmc: 4517574doi: 10.2460/ajvr.76.4.358google scholar: lookup
  66. Zeve D, Stas E, de Sousa CJ, Mannam P, Qi W, Yin X, Dubois S, Shah MS, Syverson EP, Hafner S, Karp JM, Carlone DL, Ordovas-Montanes J, Breault DT. Robust differentiation of human enteroendocrine cells from intestinal stem cells.. Nat Commun 13:261.
    pubmed: 35017529pmc: 8752608doi: 10.1038/s41467-021-27901-5google scholar: lookup
  67. Sanman LE, Chen IW, Bieber JM, Steri V, Trentesaux C, Hann B, Klein OD, Wu LF, Altschuler SJ. Transit-amplifying cells coordinate changes in intestinal epithelial cell-type composition.. Dev Cell 56:356-365.e9.
    pubmed: 33484640pmc: 7917018doi: 10.1016/j.devcel.2020.12.020google scholar: lookup
  68. Reynolds A, Wharton N, Parris A, Mitchell E, Sobolewski A, Kam C, Bigwood L, El Hadi A, Münsterberg A, Lewis M, Speakman C, Stebbings W, Wharton R, Sargen K, Tighe R, Jamieson C, Hernon J, Kapur S, Oue N, Yasui W, Williams MR. Canonical Wnt signals combined with suppressed TGFβ/BMP pathways promote renewal of the native human colonic epithelium.. Gut 63:610–621.
    pubmed: 23831735doi: 10.1136/gutjnl-2012-304067google scholar: lookup
  69. Binnerts ME, Kim KA, Bright JM, Patel SM, Tran K, Zhou M, Leung JM, Liu Y, Lomas WE 3rd, Dixon M, Hazell SA, Wagle M, Nie WS, Tomasevic N, Williams J, Zhan X, Levy MD, Funk WD, Abo A. R-Spondin1 regulates Wnt signaling by inhibiting internalization of LRP6.. Proc Natl Acad Sci USA 104:14700–14705.
    pubmed: 17804805pmc: 1965484doi: 10.1073/pnas.0702305104google scholar: lookup
  70. Sato T, van Es JH, Snippert HJ, Stange DE, Vries RG, van den Born M, Barker N, Shroyer NF, van de Wetering M, Clevers H. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts.. Nature 469:415–418.
    pubmed: 21113151doi: 10.1038/nature09637google scholar: lookup
  71. Berková L, Fazilaty H, Yang Q, Kubovčiak J, Stastna M, Hrckulak D, Vojtechova M, Dalessi T, Brügger MD, Hausmann G, Liberali P, Korinek V, Basler K, Valenta T. Terminal differentiation of villus tip enterocytes is governed by distinct Tgfβ superfamily members.. EMBO Rep 24:e56454.
    pubmed: 37493498pmc: 10481656doi: 10.15252/embr.202256454google scholar: lookup
  72. Auclair BA, Benoit YD, Rivard N, Mishina Y, Perreault N. Bone morphogenetic protein signaling is essential for terminal differentiation of the intestinal secretory cell lineage.. Gastroenterology 133:887–896.
    pubmed: 17678919doi: 10.1053/j.gastro.2007.06.066google scholar: lookup
  73. Dieterich W, Neurath MF, Zopf Y. Intestinal ex vivo organoid culture reveals altered programmed crypt stem cells in patients with celiac disease.. Sci Rep 10:3535.
    pubmed: 32103108pmc: 7044285doi: 10.1038/s41598-020-60521-5google scholar: lookup
  74. Gabriel V, Zdyrski C, Sahoo DK, Dao K, Bourgois-Mochel A, Atherly T, Martinez MN, Volpe DA, Kopper J, Allenspach K, Mochel JP. Canine intestinal organoids in a dual-chamber permeable support system.. J Vis Exp 181:e63612.

Citations

This article has been cited 3 times.
  1. Richardson LM, Gordon J, Davila C, Chamoun-Emanuelli AM, Zdyrski C, Whitfield-Cargile CM. Functional and molecular characterization of equine intestinal organoids across media conditions and intestinal segments. BMC Vet Res 2026 Jan 16;22(1).
    doi: 10.1186/s12917-025-05222-9pubmed: 41545869google scholar: lookup
  2. Stewart AS, Kopper JJ, McKinney-Aguirre C, Veerasamy B, Sahoo DK, Freund JM, Gonzalez LM. Assessment of equine intestinal epithelial junctional complexes and barrier permeability using a monolayer culture system. Front Vet Sci 2024;11:1455262.
    doi: 10.3389/fvets.2024.1455262pubmed: 39502947google scholar: lookup
  3. Kambayashi Y, Nemoto M, Ochi A, Kishi D, Ueno T, Tsujimura K, Bannai H, Kawanishi N, Ohta M, Suzuki T. Equine coronavirus infection and replication in equine intestinal enteroids. Vet Res 2024 Oct 10;55(1):135.
    doi: 10.1186/s13567-024-01381-zpubmed: 39390558google scholar: lookup
Subscribe to our newsletter
Join 99,658 horse owners like you who receive our email updates on horse health and nutrition.