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Home / Equine Research Bank / Equine Vet J / Advanced three-dimensional culture of equine intestinal epit...
Equine veterinary journal2017; 50(2); 241-248; doi: 10.1111/evj.12734

Advanced three-dimensional culture of equine intestinal epithelial stem cells.

Abstract: Intestinal epithelial stem cells are critical to epithelial repair following gastrointestinal injury. The culture of intestinal stem cells has quickly become a cornerstone of a vast number of new research endeavours that range from determining tissue viability to testing drug efficacy for humans. This study aims to describe the methods of equine stem cell culture and highlights the future benefits of these techniques for the advancement of equine medicine. Objective: To describe the isolation and culture of small intestinal stem cells into three-dimensional (3D) enteroids in horses without clinical gastrointestinal abnormalities. Methods: Descriptive study. Methods: Intestinal samples were collected by sharp dissection immediately after euthanasia. Intestinal crypts containing intestinal stem cells were dissociated from the underlying tissue layers, plated in a 3D matrix and supplemented with growth factors. After several days, resultant 3D enteroids were prepared for immunofluorescent imaging and polymerase chain reaction (PCR) analysis to detect and characterise specific cell types present. Intestinal crypts were cryopreserved immediately following collection and viability assessed. Results: Intestinal crypts were successfully cultured and matured into 3D enteroids containing a lumen and budding structures. Immunofluorescence and PCR were used to confirm the existence of stem cells and all post mitotic, mature cell types, described to exist in the horse intestinal epithelium. Previously frozen crypts were successfully cultured following a freeze-thaw cycle. Conclusions: Tissues were all derived from normal horses. Application of this technique for the study of specific disease was not performed at this time. Conclusions: The successful culture of equine intestinal crypts into 3D "mini-guts" allows for in vitro studies of the equine intestine. Additionally, these results have relevance to future development of novel therapies that harness the regenerative potential of equine intestine in horses with gastrointestinal disease (colic).
© 2017 EVJ Ltd.
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Publication Date: 2017-09-06 PubMed ID: 28792626PubMed Central: PMC5796842DOI: 10.1111/evj.12734Google Scholar: Lookup
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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 study describes the method of growing equine intestinal stem cells in a 3D format for future research purposes in veterinary medicine.

Study Overview

The research revolves around isolating and culturing small equine intestinal stem cells which are crucial for intestinal repair. These cells were grown into three-dimensional (3D) formations known as enteroids, a process that could greatly contribute to research methods and advancements in equine medicine.

Methods

  • The researchers collected intestinal samples immediately after euthanizing the horses through sharp dissection. These samples contained intestinal crypts that house the intestinal stem cells.
  • The intestinal crypts were separated from the underlying tissue layers, placed in a 3D matrix, and supplemented with growth factors. This process led to the formation of 3D enteroids (mini intestines) after several days.
  • The researchers used immunofluorescent imaging and polymerase chain reaction (PCR) analysis on the produced 3D enteroids in order to identify and classify the specific cell types found within them. They also cryopreserved some intestinal crypts immediately after collection and then assessed their viability.

Results

  • The research team successfully cultured intestinal crypts and matured them into 3D enteroids which contained a lumen and budding structures, analogous to actual intestines.
  • The presence of stem cells and all post mitotic, mature cell types, known to exist in horse’s intestinal epithelium, were confirmed through immunofluorescence and PCR.
  • The researchers also successfully cultured the previously frozen crypts, proving that the cells could survive the freezing process to be used at later times.

Conclusion

  • All tissues used in the study were derived from healthy horses. The team did not utilize this technique to study any specific diseases during this study.
  • The successful culture of equine intestinal crypts into 3D “mini-guts” opens up the possibility for in vitro studies of the equine intestine, presenting new opportunities for research.
  • This breakthrough might also have a significant impact on the development of novel therapies that leverage the regenerative potential of the equine intestine to treat horses with gastrointestinal diseases such as colic.

Cite This Article

APA
Stewart AS, Freund JM, Gonzalez LM. (2017). Advanced three-dimensional culture of equine intestinal epithelial stem cells. Equine Vet J, 50(2), 241-248. https://doi.org/10.1111/evj.12734

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 50
Issue: 2
Pages: 241-248

Researcher Affiliations

Stewart, A Stieler
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.
Freund, J M
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.
Gonzalez, L M
  • Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA.

MeSH Terms

  • Animals
  • Cell Culture Techniques / veterinary
  • Cell Differentiation
  • Female
  • Horses
  • Intestinal Mucosa / cytology
  • Male
  • Models, Biological
  • Stem Cells / physiology

Grant Funding

  • K01 OD019911 / NIH HHS
  • T32 OD011130 / NIH HHS

Conflict of Interest Statement

. No competing interests have been declared.

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Citations

This article has been cited 15 times.
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