Background and Purpose: Epidemiological studies suggest that maternal exposures to environmental chemicals may be linked to spontaneous preterm birth. Mechanistic studies are needed to verify these findings; however, existing models (rodents, human explant placentas or traditional in vitro studies) do not resemble the human feto-maternal interfaces that function as a major barrier and immune responder to environmental chemicals. The four-cell human Feto-Maternal interface Organ-On-Chip (FMi-OOC) model was recently developed for studies of chemical transport and effects across this barrier. Methods: Four environmental compounds with suggestive epidemiological evidence for preterm birth outcomes were used in these studies: dichlorodiphenyltrichloroethane (DDT), bisphenol A (BPA), 2,2’4,4’-tetrabromodiphenyl ether (PBDE-47), and perfluorooctanoic acid (PFOA). First, we tested cytotoxicity of these compounds in concentration response (1 to 300 μM) in four human immortalized fetal-maternal cell types (maternal decidua, and fetal membranes’ chorion trophoblast, amnion mesenchymal, and amnion epithelial cells). Second, we selected non-cytotoxic concentrations (from 1 to 100 μM depending on a compound) for testing in FMi-OOC devices that were made using a soft-lithography fabrication technique and plasma bonding process. Each FMi-OOC is composed of four concentric ring-shaped chambers connected by arrays of microchannels. Each chamber is seeded with one fetalmaternal cell type in order of their position in the human FMi to mimic the in utero environment. Test compounds were added to the maternal decidua chamber and chemical transport, cell viability, and proinflammatory cytokine (IL-6, IL-8, GMCSF, and TNF-α) response from each cell type was determined. Results: Dose-response studies using traditional 2D cultures enabled experimentally-informed selection of the concentration ranges, a necessary step for subsequent studies in a more complex and lower throughput FMi-OOC model. In FMi-OOC, the analytical chemistry data showed that the majority of each test compound remained in the maternal chamber where it was added. Each compound propagated only to the minimal extent to the fetal chambers. All test compounds had no cytotoxic effects on the maternal decidua cells. However, they exhibited some cytotoxic effects in the fetal chambers, most of these were observed in the chorion trophoblast cells. In our study, treatment-associated effects on proinflammatory cytokine production were observed with all test compounds, most notable effects were found with DDT and the most responsive cell type across all tested substances was fetal chorion trophoblast. Conclusions: These studies provide valuable mechanistic support to the epidemiological observations that maternal exposure of DDT, BPA, PBDE-47 and PFOA may lead to a fetal inflammatory response, as evidenced by cytokine increase, that can trigger the pathways of preterm birth. We report the development of a microphysiological system (FMi-OOC) that could be used for studies of maternal environmental chemical exposures and their potential effects on the fetal tissues.