title: An inter-laboratory study to evaluate the effects of medium composition on the differentiation and barrier function of Caco-2 cell lines.
abstract: Differentiated human intestinal Caco-2 cells are frequently used in toxicology and pharmacology as in vitro models for studies on intestinal barrier functions. Since several discrepancies exist among the different lines and clones of Caco-2 cells, comparison of the results obtained and optimisation of models for use for regulatory purposes are particularly difficult, especially with respect to culture conditions and morphological and biochemical parameters. An inter-laboratory study has been performed on the parental cell line and on three clonal Caco-2 cell lines, with the aim of standardising the culture conditions and identifying the best cell line with respect to parameters relevant to barrier integrity, namely, trans-epithelial electrical resistance (TEER) and mannitol passage, and of epithelial differentiation (alkaline phosphatase activity). Comparison of the cell lines maintained in traditional serum-supplemented culture medium or in defined medium, containing insulin, transferrin, selenium and lipids, showed that parameter performance was better and more reproducible with the traditional medium. The maintenance of the cell lines for 15 days in culture was found to be sufficient for the development of barrier properties, but not for full epithelial differentiation. Caco-2/TC7 cells performed better than the other three cell lines, both in terms of reproducibility and performance, exhibiting low TEER and mannitol passage, and high alkaline phosphatase activity.
authors.affiliation: Istituto di Neurobiologia e Medicina Molecolare, CNR, Rome, Italy.
authors.forename: Flavia
authors.initials: F
authors.lastname: Zucco
authors.forename: Anne-Françoise
authors.initials: AF
authors.lastname: Batto
authors.forename: Giovanna
authors.initials: G
authors.lastname: Bises
authors.forename: Jean
authors.initials: J
authors.lastname: Chambaz
authors.forename: Arianna
authors.initials: A
authors.lastname: Chiusolo
authors.forename: Rosa
authors.initials: R
authors.lastname: Consalvo
authors.forename: Heide
authors.initials: H
authors.lastname: Cross
authors.forename: Gianni
authors.initials: G
authors.lastname: Dal Negro
authors.forename: Isabella
authors.initials: I
authors.lastname: de Angelis
authors.forename: Gérard
authors.initials: G
authors.lastname: Fabre
authors.forename: François
authors.initials: F
authors.lastname: Guillou
authors.forename: Sebastian
authors.initials: S
authors.lastname: Hoffman
authors.forename: Loïc
authors.initials: L
authors.lastname: Laplanche
authors.forename: Etienne
authors.initials: E
authors.lastname: Morel
authors.forename: Martine
authors.initials: M
authors.lastname: Pinçon-Raymond
authors.forename: Pilar
authors.initials: P
authors.lastname: Prieto
authors.forename: Laura
authors.initials: L
authors.lastname: Turco
authors.forename: Giulia
authors.initials: G
authors.lastname: Ranaldi
authors.forename: Monique
authors.initials: M
authors.lastname: Rousset
authors.forename: Yula
authors.initials: Y
authors.lastname: Sambuy
authors.forename: Maria Laura
authors.initials: ML
authors.lastname: Scarino
authors.forename: François
authors.initials: F
authors.lastname: Torreilles
authors.forename: Annalaura
authors.initials: A
authors.lastname: Stammati
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000704:Analysis of Variance; D015415:Biomarkers / Q000032:analysis; D018938:Caco-2 Cells / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D017097:Electric Impedance; D006801:Humans; D008353:Mannitol / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D015415:Biomarkers; D003470:Culture Media; D008353:Mannitol; D000469:Alkaline Phosphatase