Biorelevant media resistant co-culture model mimicking permeability of human intestine

• Published in: International journal of pharmaceutics Vol. 481; no. 1-2; pp. 27 - 36
• Main Authors: Antoine, Delphine, Pellequer, Yann, Tempesta, Camille, Lorscheidt, Stefan, Kettel, Bernadette, Tamaddon, Lana, Jannin, Vincent, Demarne, Frédéric, Lamprecht, Alf, Béduneau, Arnaud
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.03.2015
• Subjects: ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism; Biological Transport; Biorelevant media; Caco-2 Cells; Cell Survival; Co-culture; Coculture Techniques; HT29 Cells; Humans; Intestinal Absorption; Intestinal Secretions; Intestines - metabolism; Mucus; Mucus - metabolism; Permeability; Propranolol - pharmacology; P-gp; Biorelevant media; Caco-2 cells; FaSSIF; Mucus; MTT; LY; TB; HCl; MTX; BSA; LDH; Papp; ANOVA; FeSSIF; HEPES; FCS; NACC; TEER; HBSS; Co-culture; Intestinal absorption; DPBS
• ISSN: 0378-5173; 1873-3476
• DOI: 10.1016/j.ijpharm.2015.01.028

[Display omitted] 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.