A novel method for isolation and culture of primary swine gastric epithelial cells
Digital
- Autores:
-
Bautista-Amorocho, Henry
Silva-Sayago, Jorge Alexander
Goyeneche-Patino, Diego A.
Pérez-Cala, Tania Liseth
Macías-Gómez, Fabio
Arango-Viana, Juan Carlos
Martínez, Alonso
- Tipo de recurso:
- Article of journal
- Fecha de publicación:
- 2021
- Institución:
- Universidad de Santander
- Repositorio:
- Repositorio Universidad de Santander
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.udes.edu.co:001/6199
- Acceso en línea:
- https://doi.org/10.1186/s12860-020-00341-7
https://repositorio.udes.edu.co/handle/001/6199
- Palabra clave:
- Cell culture
Swine gastric epithelium
Tissue engineering
Animal models
Biotechnology
- Rights
- openAccess
- License
- © The Author(s). 2021
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A novel method for isolation and culture of primary swine gastric epithelial cells |
| title |
A novel method for isolation and culture of primary swine gastric epithelial cells |
| spellingShingle |
A novel method for isolation and culture of primary swine gastric epithelial cells Cell culture Swine gastric epithelium Tissue engineering Animal models Biotechnology |
| title_short |
A novel method for isolation and culture of primary swine gastric epithelial cells |
| title_full |
A novel method for isolation and culture of primary swine gastric epithelial cells |
| title_fullStr |
A novel method for isolation and culture of primary swine gastric epithelial cells |
| title_full_unstemmed |
A novel method for isolation and culture of primary swine gastric epithelial cells |
| title_sort |
A novel method for isolation and culture of primary swine gastric epithelial cells |
| dc.creator.fl_str_mv |
Bautista-Amorocho, Henry Silva-Sayago, Jorge Alexander Goyeneche-Patino, Diego A. Pérez-Cala, Tania Liseth Macías-Gómez, Fabio Arango-Viana, Juan Carlos Martínez, Alonso |
| dc.contributor.author.none.fl_str_mv |
Bautista-Amorocho, Henry Silva-Sayago, Jorge Alexander Goyeneche-Patino, Diego A. Pérez-Cala, Tania Liseth Macías-Gómez, Fabio Arango-Viana, Juan Carlos Martínez, Alonso |
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CliniUDES |
| dc.subject.proposal.eng.fl_str_mv |
Cell culture Swine gastric epithelium Tissue engineering Animal models Biotechnology |
| topic |
Cell culture Swine gastric epithelium Tissue engineering Animal models Biotechnology |
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Digital |
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2021 |
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2021-01-06 |
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2022-03-01T16:25:53Z |
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2022-03-01T16:25:53Z |
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https://doi.org/10.1186/s12860-020-00341-7 |
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https://repositorio.udes.edu.co/handle/001/6199 |
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https://doi.org/10.1186/s12860-020-00341-7 https://repositorio.udes.edu.co/handle/001/6199 |
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eng |
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eng |
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Bautista-Amorocho, H., Silva-Sayago, J.A., Goyeneche-Patino, D.A. et al. A novel method for isolation and culture of primary swine gastric epithelial cells. BMC Mol and Cell Biol 22, 1 (2021). https://doi.org/10.1186/s12860-020-00341-7 |
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Scopus |
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BMC Molecular and Cell Biology |
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© The Author(s). 2021 |
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info:eu-repo/semantics/openAccess |
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Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0) |
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https://creativecommons.org/licenses/by-nc/4.0/ |
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© The Author(s). 2021 Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0) https://creativecommons.org/licenses/by-nc/4.0/ http://purl.org/coar/access_right/c_abf2 |
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openAccess |
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13 p |
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application/pdf |
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Reino Unido |
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https://bmcmolcellbiol.biomedcentral.com/track/pdf/10.1186/s12860-020-00341-7.pdf |
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Universidad de Santander |
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Bautista-Amorocho, Henryd9a4964c-5a30-4208-8e21-c05d2e03f8ab-1Silva-Sayago, Jorge Alexandere66f05d9-3dd3-45e7-9727-a0180eea59b5-1Goyeneche-Patino, Diego A.fcf7ce5b-de44-4ffa-aaea-502fa3ad564e-1Pérez-Cala, Tania Liseth0ccfdfc3-f2f5-4f05-8694-ea6da5631ae1-1Macías-Gómez, Fabio860cc931-035c-489f-b8b1-327c5709bc70-1Arango-Viana, Juan Carlos90a1676c-4b45-4a35-9bcf-846b26171e37-1Martínez, Alonsocef03e01-d11f-4351-adc7-accf410669e2-1CliniUDES2022-03-01T16:25:53Z2022-03-01T16:25:53Z2021-01-06DigitalBackground: Culturing primary epithelial cells has a major advantage over tumor-derived or immortalized cell lines as long as their functional phenotype and genetic makeup are mainly maintained. The swine model has shown to be helpful and reliable when used as a surrogate model for human diseases. Several porcine cell lines have been established based on a variety of tissues, which have shown to extensively contribute to the current understanding of several pathologies, especially cancer. However, protocols for the isolation and culture of swine gastric epithelial cells that preserve cell phenotype are rather limited. We aimed to develop a new method for establishing a primary epithelial cell culture from the fundic gland region of the pig stomach. Results: Mechanical and enzymatic dissociation of gastric tissue was possible by combining collagenase type I and dispase II, protease inhibitors and antioxidants, which allowed the isolation of epithelial cells from the porcine fundic glands showing cell viability > 90% during the incubation period. Gastric epithelial cells cultured in RPMI 1640, DMEM-HG and DMEM/F12 media did not contribute enough to cell adhesion, cluster formation and cell proliferation. By contrast, William’s E medium supplemented with growth factors supports confluency and proliferation of a pure epithelial cell monolayer after 10 days of incubation at 37 °C, 5% CO2. Mucin-producing cell phenotype of primary isolates was confirmed by PAS staining, MUC1 by immunohistochemistry, as well as the expression of MUC1 and MUC20 genes by RT-PCR and cDNA sequencing. Swine gastric epithelial cells also showed origin-specific markers such as cytokeratin cocktail (AE1/AE3) and cytokeratin 18 (CK-18) using immunohistochemical and immunofluorescence methods, respectively. Conclusions: A new method was successfully established for the isolation of primary gastric epithelial cells from the fundic gland zone through a swine model based on a combination of tissue-specific proteases, protease inhibitors and antioxidants after mechanical cell dissociation. The formulation of William’s E medium with growth factors for epithelial cells contributes to cell adhesion and preserves functional primary cells phenotype, which is confirmed by mucin production and expression of typical epithelial markers over time.Ciencias Médicas y de la Salud13 papplication/pdfhttps://doi.org/10.1186/s12860-020-00341-7https://repositorio.udes.edu.co/handle/001/6199engReino Unido131122Bautista-Amorocho, H., Silva-Sayago, J.A., Goyeneche-Patino, D.A. et al. A novel method for isolation and culture of primary swine gastric epithelial cells. BMC Mol and Cell Biol 22, 1 (2021). https://doi.org/10.1186/s12860-020-00341-7ScopusBMC Molecular and Cell Biology© The Author(s). 2021info:eu-repo/semantics/openAccessAtribución-NoComercial 4.0 Internacional (CC BY-NC 4.0)https://creativecommons.org/licenses/by-nc/4.0/http://purl.org/coar/access_right/c_abf2https://bmcmolcellbiol.biomedcentral.com/track/pdf/10.1186/s12860-020-00341-7.pdfCell cultureSwine gastric epitheliumTissue engineeringAnimal modelsBiotechnologyA novel method for isolation and culture of primary swine gastric epithelial cellsArtículo de revistahttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85Todas las AudienciasPublicationORIGINALA novel method for isolation and culture of primary swine gastric epithelial cells.pdfA novel method for isolation and culture of primary swine gastric epithelial cells.pdfapplication/pdf231198https://repositorio.udes.edu.co/bitstreams/ce9bfa46-daeb-46a7-bd18-e79541de85dc/download7a6e57638961968b9237d80a4ad9abb5MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-859https://repositorio.udes.edu.co/bitstreams/739a647f-9805-4ad5-92d1-20dcfbc1f546/download38d94cf55aa1bf2dac1a736ac45c881cMD52TEXTA novel method for isolation and culture of primary swine gastric epithelial cells.pdf.txtA novel method for isolation and culture of primary swine gastric epithelial cells.pdf.txtExtracted texttext/plain5https://repositorio.udes.edu.co/bitstreams/121fc5dd-4997-4209-81a6-191415d7235e/download5dbe86c1111d64f45ba435df98fdc825MD53THUMBNAILA novel method for isolation and culture of primary swine gastric epithelial cells.pdf.jpgA novel method for isolation and culture of primary swine gastric epithelial cells.pdf.jpgGenerated Thumbnailimage/jpeg12989https://repositorio.udes.edu.co/bitstreams/8d363f37-6d9e-4fcd-957d-770d21513c52/download4cf14b1deebdae4adbc147d1d82f37d1MD54001/6199oai:repositorio.udes.edu.co:001/61992023-10-10 09:50:23.311https://creativecommons.org/licenses/by-nc/4.0/© The Author(s). 2021https://repositorio.udes.edu.coRepositorio Universidad de Santandersoporte@metabiblioteca.comTGljZW5jaWEgZGUgUHVibGljYWNpw7NuIFVERVMKRGlyZWN0cmljZXMgZGUgVVNPIHkgQUNDRVNPCgo= |