title: Hepatocytes cocultured with Sertoli cells in bioreactor favors Sertoli barrier tightness in rat.
abstract: The lack of a reliable in vitro system to assess reprotoxicity is an emerging problem in the context of European law for Registration, Evaluation, Authorization and Restriction of Chemicals (REACH, 2007), as it requires a reduction in animal utilization for testing. Furthermore, in vitro reprotoxicological tests would be more relevant and greatly improved by integrating both hepatic metabolism and the blood-testis barrier. Here, we took advantage of an integrated insert in a dynamic microfluidic platform (IIDMP) to co-cultivate hepatocytes in biochip and Sertoli cells in the bicameral chamber. This microfluidic tool has been previously demonstrated to be helpful in cell function and/or quality improvement. We demonstrate that permeability of the Sertoli barrier is reduced by dynamic coculture in our system. Exometabolomics analysis reveals that interactions between hepatocytes and Sertoli cells may have been mediated by the polyamines increase and/or mid-chain fatty acid decrease in the circulating medium. These metabolic changes may be involved in permeability reduction by contributing to modifying junction protein quantity and localization. The present study gives an example of IIDMP as an in vitro partitioning/transport model for cell culture and toxicological testing. Further, based on both our previous results using an intestinal-hepatic cell coculture and the present study, IIDMP seems to be well-suited for (i) assessing the dose-response effect of chemicals within the rodent or human male reproductive tract, and (ii) improving the quality of reprotoxicological assays by including hepatic metabolism. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Biomécanique et Bioingénierie, Centre de recherche Royallieu, Sorbonne universités, Université de Technologie de Compiègne, CNRS, UMR 7338, Compiègne cedex, France.|INSERM U1193, Physiopathogénèse et Traitement des Maladies du Foie, Hôpital Paul Brousse, bâtiment Lavoisier, Villejuif Cedex, France.
authors.forename: P
authors.initials: P
authors.lastname: Zeller
authors.affiliation: Biomécanique et Bioingénierie, Centre de recherche Royallieu, Sorbonne universités, Université de Technologie de Compiègne, CNRS, UMR 7338, Compiègne cedex, France.|Institut de Radioprotection et de Sûreté Nucléaire (IRSN)PRP-HOM, SRBE, LRTOX, Fontenay-aux-Roses Cedex, France.
authors.forename: A
authors.initials: A
authors.lastname: Legendre
authors.affiliation: INSERM U1016, Plate-forme génomique, institut Cochin, 22 rue Méchain, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Jacques
authors.affiliation: Biomécanique et Bioingénierie, Centre de recherche Royallieu, Sorbonne universités, Université de Technologie de Compiègne, CNRS, UMR 7338, Compiègne cedex, France.
authors.forename: M J
authors.initials: MJ
authors.lastname: Fleury
authors.affiliation: Institute of Plant Sciences Paris-Saclay (IPS2), UMR 9213/UMR1403, CNRS, INRA, Saclay Plant Sciences, Université Paris-Sud, Université d'Evry, Université Paris-Diderot, Sorbonne Paris-Cité, France.
authors.forename: F
authors.initials: F
authors.lastname: Gilard
authors.affiliation: Research School of Biology, ANU College of Medicine, Biology and Environment, Australian National University, Canberra ACT, Australia.
authors.forename: G
authors.initials: G
authors.lastname: Tcherkez
authors.affiliation: Biomécanique et Bioingénierie, Centre de recherche Royallieu, Sorbonne universités, Université de Technologie de Compiègne, CNRS, UMR 7338, Compiègne cedex, France.|CNRS-LIMMS-UMI 2820, Institute of Industrial Science, University of Tokyo, Meguro ku, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Leclerc
mesh_terms: D023401:Animal Use Alternatives; D000818:Animals; D019149:Bioreactors; D001814:Blood-Testis Barrier; D018920:Coculture Techniques; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D044085:Microfluidics; D010539:Permeability; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019108:Tight Junctions / Q000187:drug effects* / Q000378:metabolism
keywords: Sertoli cell epithelium permeability; coculture; exometabolomics; microfluidic; reprotoxicity; tight junctions