Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells

Intestinal epithelial cell culture is important for biological, functional, and immunological studies. Since enterocytes have a short in vivo life span due to anoikis, we aimed to establish a novel and reproducible method to prolong the survival of mouse and human cells. Cells were isolated followin...

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Autores principales: Di Claudio, Fiorella, Muglia, Cecilia Isabel, Smaldini, Paola Lorena, Orsini Delgado, María Lucía, Trejo, Fernando Miguel, Grigera, José Raúl, Docena, Guillermo Horacio
Formato: Articulo Preprint
Lenguaje:Inglés
Publicado: 2017
Materias:
Química
Biología
Enterocyte
Cell culture
Pprimary culture
Collagen membrane
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/127382
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-127382
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Química
Biología
Enterocyte
Cell culture
Pprimary culture
Collagen membrane
spellingShingle Química
Biología
Enterocyte
Cell culture
Pprimary culture
Collagen membrane
Di Claudio, Fiorella
Muglia, Cecilia Isabel
Smaldini, Paola Lorena
Orsini Delgado, María Lucía
Trejo, Fernando Miguel
Grigera, José Raúl
Docena, Guillermo Horacio
Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells
topic_facet Química
Biología
Enterocyte
Cell culture
Pprimary culture
Collagen membrane
description Intestinal epithelial cell culture is important for biological, functional, and immunological studies. Since enterocytes have a short in vivo life span due to anoikis, we aimed to establish a novel and reproducible method to prolong the survival of mouse and human cells. Cells were isolated following a standard procedure, and cultured on ordered-cow's collagen membranes. A prolonged cell life span was achieved; cells covered the complete surface of bio-membranes and showed a classical enterocyte morphology with high expression of enzymes supporting the possibility of cryopreservation. Apoptosis was dramatically reduced and cultured enterocytes expressed cytokeratin and LGR5 (low frequency). Cells exposed to LPS or flagellin showed the induction of TLR4 and TLR5 expression and a functional phenotype upon exposure to the probiotic Bifidobacterium bifidum or the pathogenic Clostridium difficile. The secretion of the homeostatic (IL-25 and TSLP), inhibitory (IL-10 and TGF-β), or pro-inflammatory mediators (IL-1β and TNF) were induced. In conclusion, this novel protocol using cow's collagen-ordered membrane provides a simple and reproducible method to maintain intestinal epithelial cells functional for cell-microorganism interaction studies and stem cell expansion.
format Articulo
Preprint
author Di Claudio, Fiorella
Muglia, Cecilia Isabel
Smaldini, Paola Lorena
Orsini Delgado, María Lucía
Trejo, Fernando Miguel
Grigera, José Raúl
Docena, Guillermo Horacio
author_facet Di Claudio, Fiorella
Muglia, Cecilia Isabel
Smaldini, Paola Lorena
Orsini Delgado, María Lucía
Trejo, Fernando Miguel
Grigera, José Raúl
Docena, Guillermo Horacio
author_sort Di Claudio, Fiorella
title Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells
title_short Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells
title_full Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells
title_fullStr Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells
title_full_unstemmed Use of a collagen membrane to enhance the survival of primary intestinal epithelial cells
title_sort use of a collagen membrane to enhance the survival of primary intestinal epithelial cells
publishDate 2017
url http://sedici.unlp.edu.ar/handle/10915/127382
work_keys_str_mv AT diclaudiofiorella useofacollagenmembranetoenhancethesurvivalofprimaryintestinalepithelialcells
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bdutipo_str Repositorios
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