title: Biorelevant media resistant co-culture model mimicking permeability of human intestine.
abstract: Cell culture models are currently used to predict absorption pattern of new compounds and formulations in the human gastro-intestinal tract (GIT). One major drawback is the lack of relevant apical incubation fluids allowing mimicking luminal conditions in the GIT. Here, we suggest a culture model compatible with biorelevant media, namely Fasted State Simulated Intestinal Fluid (FaSSIF) and Fed State Simulated Intestinal Fluid (FeSSIF). Co-culture was set up from Caco-2 and mucus-secreting HT29-MTX cells using an original seeding procedure. Viability and cytotoxicity assays were performed following incubation of FeSSIF and FaSSIF with co-culture. Influence of biorelevant fluids on paracellular permeability or transporter proteins were also evaluated. Results were compared with Caco-2 and HT29-MTX monocultures. While Caco-2 viability was strongly affected with FeSSIF, no toxic effect was detected for the co-cultures in terms of viability and lactate dehydrogenase release. The addition of FeSSIF to the basolateral compartment of the co-culture induced cytotoxic effects which suggested the apical mucus barrier being cell protective. In contrast to FeSSIF, FaSSIF induced a slight increase of the paracellular transport and both tested media inhibited partially the P-gp-mediated efflux in the co-culture. Additionally, the absorptive transport of propranolol hydrochloride, a lipophilic β-blocker, was strongly affected by biorelevant fluids. This study demonstrated the compatibility of the Caco-2/HT29-MTX model with some of the current biorelevant media. Combining biorelevant intestinal fluids with features such as mucus secretion, adjustable paracellular and P-gp mediated transports, is a step forward to more realistic in-vitro models of the human intestine.
authors.affiliation: EA 4267 Fonctions et Dysfonctions Epitheliales, University of Franche-Comté, Besançon, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Antoine
authors.affiliation: EA 4267 Fonctions et Dysfonctions Epitheliales, University of Franche-Comté, Besançon, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Pellequer
authors.affiliation: EA 4267 Fonctions et Dysfonctions Epitheliales, University of Franche-Comté, Besançon, France.
authors.forename: Camille
authors.initials: C
authors.lastname: Tempesta
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmaceutics, University of Bonn, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Lorscheidt
authors.affiliation: Laboratory of Pharmaceutical Technology and Biopharmaceutics, University of Bonn, Germany.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Kettel
authors.affiliation: EA 4267 Fonctions et Dysfonctions Epitheliales, University of Franche-Comté, Besançon, France.
authors.forename: Lana
authors.initials: L
authors.lastname: Tamaddon
authors.affiliation: Gattefossé SAS, 36 chemin de Genas, 69804 Saint-Priest Cedex, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Jannin
authors.affiliation: Gattefossé SAS, 36 chemin de Genas, 69804 Saint-Priest Cedex, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Demarne
authors.affiliation: EA 4267 Fonctions et Dysfonctions Epitheliales, University of Franche-Comté, Besançon, France; Laboratory of Pharmaceutical Technology and Biopharmaceutics, University of Bonn, Germany.
authors.forename: Alf
authors.initials: A
authors.lastname: Lamprecht
authors.affiliation: EA 4267 Fonctions et Dysfonctions Epitheliales, University of Franche-Comté, Besançon, France. Electronic address: arnaud.beduneau@univ-fcomte.fr.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Béduneau
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D001692:Biological Transport; D018938:Caco-2 Cells; D002470:Cell Survival; D018920:Coculture Techniques; D019073:HT29 Cells; D006801:Humans; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000378:metabolism*; D007419:Intestinal Secretions; D009093:Mucus / Q000378:metabolism; D010539:Permeability; D011433:Propranolol / Q000494:pharmacology
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D011433:Propranolol
keywords: Biorelevant media; Caco-2 cells; Co-culture; Intestinal absorption; Mucus