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A novel method for isolation and culture of primary swine gastric epithelial cells

dc.contributor.authorBautista-Amorocho, Henry
dc.contributor.authorSilva-Sayago, Jorge Alexander
dc.contributor.authorGoyeneche-Patino, Diego A.
dc.contributor.authorPérez-Cala, Tania Liseth
dc.contributor.authorMacías-Gómez, Fabio
dc.contributor.authorArango-Viana, Juan Carlos
dc.contributor.authorMartínez, Alonso
dc.contributor.researchgroupCliniUDESspa
dc.date.accessioned2022-03-01T16:25:53Z
dc.date.available2022-03-01T16:25:53Z
dc.date.issued2021-01-06
dc.descriptionDigitalspa
dc.description.abstractBackground: 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.eng
dc.description.researchareaCiencias Médicas y de la Saludspa
dc.format.extent13 pspa
dc.format.mimetypeapplication/pdfspa
dc.identifier.doihttps://doi.org/10.1186/s12860-020-00341-7
dc.identifier.urihttps://repositorio.udes.edu.co/handle/001/6199
dc.language.isoengspa
dc.publisher.placeReino Unidospa
dc.relation.citationendpage13spa
dc.relation.citationissue1spa
dc.relation.citationstartpage1spa
dc.relation.citationvolume22spa
dc.relation.citesBautista-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
dc.relation.indexedScopusspa
dc.relation.ispartofjournalBMC Molecular and Cell Biologyspa
dc.rights© The Author(s). 2021spa
dc.rights.accessrightsinfo:eu-repo/semantics/openAccessspa
dc.rights.creativecommonsAtribución-NoComercial 4.0 Internacional (CC BY-NC 4.0)spa
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/spa
dc.sourcehttps://bmcmolcellbiol.biomedcentral.com/track/pdf/10.1186/s12860-020-00341-7.pdfspa
dc.subject.proposalCell cultureeng
dc.subject.proposalSwine gastric epitheliumeng
dc.subject.proposalTissue engineeringeng
dc.subject.proposalAnimal modelseng
dc.subject.proposalBiotechnologyeng
dc.titleA novel method for isolation and culture of primary swine gastric epithelial cellsspa
dc.typeArtículo de revistaspa
dc.type.coarhttp://purl.org/coar/resource_type/c_6501spa
dc.type.contentTextspa
dc.type.driverinfo:eu-repo/semantics/articlespa
dc.type.redcolhttp://purl.org/redcol/resource_type/ARTspa
dc.type.versioninfo:eu-repo/semantics/publishedVersionspa
dcterms.audienceTodas las Audienciasspa
dspace.entity.typePublication
oaire.accessrightshttp://purl.org/coar/access_right/c_abf2spa
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85spa
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