New model of human placental co-culture to evaluate the effects of food contaminants on placental barrier

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Leconte, Camille | Gomez, Camille | Nogues, Perrine | dos Santos, Esther | Dieudonne, Marie-Noëlle | Couturier-Tarrade, Anne

Edité par CCSD -

The placenta, located at the interface between the maternal and the fetal sides, playsa crucial role in nutrient exchanges, endocrine function and immunity. This organ issensitive to maternal environment (nutrition, metabolism, pollutants…), which candisrupt placental functions leading to disturbance of fetal development, fetal growthand long-term effects on offspring phenotype. To limit the use of animal experimentsto study the effects of food contaminants or other molecules on the placental function,it seems crucial to develop cellular models to evaluate their transfer and their effectson placental endocrine function.Our aim was to develop a method of co-culture using human term placenta, to beclose to in vivo placental barrier, including mesenchyme and trophoblastic cells, in atranswell® system.The method to isolate and purify mesenchyme cells from human placenta wasdeveloped using enzymatic digestion associated to percoll gradient. Cells werecharacterized by immunocytochemistry and cell concentration was adjustedaccording to cell viability using LDH measurement in the media. Layer permeabilitywas assessed by a quantification of fluorescent sodium fluorescein molecule (Na-Flu)in both transwell® compartments. Trophoblastic cells were isolated from the humanplacenta as described Kliman et al. 1986 and the timing of trophoblastic cells addingto mesenchyme cells on the transwell® was determined. LDH and Na-Fluconcentrations were measured associated to the production of the human ChorionicGonadotropin, an hormone which reflects the functionality of trophoblastic cells.Establishment of this co-culture system allows us to investigate the effects of foodadditives containing nanoparticles on placental barrier. Work is ongoing to explorethe placental transfer and endocrine function using gold-nanoparticles at differentsizes and doses.

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