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Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase
The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the prese...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Associação Brasileira de Divulgação Científica
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5441283/ https://www.ncbi.nlm.nih.gov/pubmed/28423120 http://dx.doi.org/10.1590/1414-431X20175831 |
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author | Ren, H.J. Zhang, C.L. Liu, R.D. Li, N. Li, X.G. Xue, H.K. Guo, Y. Wang, Z.Q. Cui, J. Ming, L. |
author_facet | Ren, H.J. Zhang, C.L. Liu, R.D. Li, N. Li, X.G. Xue, H.K. Guo, Y. Wang, Z.Q. Cui, J. Ming, L. |
author_sort | Ren, H.J. |
collection | PubMed |
description | The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology. |
format | Online Article Text |
id | pubmed-5441283 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Associação Brasileira de Divulgação Científica |
record_format | MEDLINE/PubMed |
spelling | pubmed-54412832017-06-05 Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase Ren, H.J. Zhang, C.L. Liu, R.D. Li, N. Li, X.G. Xue, H.K. Guo, Y. Wang, Z.Q. Cui, J. Ming, L. Braz J Med Biol Res Biomedical Sciences The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology. Associação Brasileira de Divulgação Científica 2017-04-13 /pmc/articles/PMC5441283/ /pubmed/28423120 http://dx.doi.org/10.1590/1414-431X20175831 Text en http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Biomedical Sciences Ren, H.J. Zhang, C.L. Liu, R.D. Li, N. Li, X.G. Xue, H.K. Guo, Y. Wang, Z.Q. Cui, J. Ming, L. Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase |
title | Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase |
title_full | Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase |
title_fullStr | Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase |
title_full_unstemmed | Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase |
title_short | Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase |
title_sort | primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type i collagenase and hyaluronidase |
topic | Biomedical Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5441283/ https://www.ncbi.nlm.nih.gov/pubmed/28423120 http://dx.doi.org/10.1590/1414-431X20175831 |
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