A Unique Co-culture Model for Fundamental and Applied Studies of Human Fetoplacental Steroidogenesis and Interference by Environmental Chemicals

Abstract

Background: Experimental tools for studying the complex steroidogenic interactions that occur between placenta and fetus during human pregnancy are extremely limited.Objectives: We aimed to develop a co-culture model to study steroidogenesis by the human fetoplacental unit and its disruption by exposure to environmental contaminants.Methods: We cultured BeWo human choriocarcinoma cells, representing the villous cytotrophoblast, and H295R human adrenocortical carcinoma cells, representing the fetal unit, in a carefully adapted co-culture medium. We placed H295R cells in 24-well plates and BeWo cells on transwell inserts with or without pesticide treatment (atrazine or prochloraz) and assessed CYP19 activity and hormonal production after 24 hr of co-culture.Results: The co-culture exhibited the steroidogenic profile of the fetoplacental unit, allowing a synergistic production of estradiol and estriol (but not of estrone) of 133.3 ± 11.3 pg/mL and 440.8 ± 44.0 pg/mL, respectively. Atrazine and prochloraz had cell-type specific effects on CYP19 activity and estrogen production in co-culture. Atrazine induced CYP19 activity and estrogen production in H295R cells only, but did not affect overall estrogen production in co-culture, whereas prochloraz inhibited CYP19 activity exclusively in BeWo cells and reduced estrogen production in co-culture by almost 90%. In contrast, prochloraz did not affect estradiol or estrone production in BeWo cells in monoculture. These differential effects underline the relevance of our co-culture approach to model fetoplacental steroidogenesis.Conclusions: The co-culture of H295R and BeWo cells creates a unique in vitro model to reproduce the steroidogenic cooperation between fetus and placenta during pregnancy and can be used to study the endocrine-disrupting effects of environmental chemicals.Citation: Hudon Thibeault AA, Deroy K, Vaillancourt C, Sanderson JT. 2014. A unique co-culture model for fundamental and applied studies of human fetoplacental steroidogenesis and interference by environmental chemicals. Environ Health Perspect 122:371–377; http://dx.doi.org/10.1289/ehp.1307518

Effects of Neonicotinoids on Promoter-Specific Expression and Activity of Aromatase (CYP19) in Human Adrenocortical Carcinoma (H295R) and Primary Umbilical Vein Endothelial (HUVEC) Cells

“…Effects on aromatase and consequences for human health are less well understood. It has been demonstrated that exposure to atrazine, a widely used herbicide, induces aromatase expression and estrogen biosynthesis in certain human cell lines (Sanderson et al, 2001(Sanderson et al, , 2002Thibeault et al, 2014), but the effects of chemicals on the promoter-and tissue-specific expression of aromatase are largely unknown. In placenta, CYP19 is expressed mainly via the placenta-specific promoter I.1, and the estrogens produced are essential for proper development of placenta and fetus (Albrecht and Pepe, 2010;Bukovsky et al, 2003).…”

Effects of long-term endocrine disrupting compound exposure on Macaca mulatta embryonic stem cells

“…Two additional stress-related CPs, HIF1A signaling and UVA-induced MAPK signaling, were affected. For comparison, past studies reported that ATR at 60 nM inhibits mouse T-cell proliferation [57], whereas 30 μM ATM does not inhibit proliferation but induces CYP19 activity in human placental cells [43]. …”

“…The concentration of ATR used was ~4-fold higher than the maximum concentration reported in one study in drinking water [36], and one study of occupational human serum level (up to 245 nM [42]). The ATR concentration used affects placenta cell gene expression in vitro [43] and is much lower than the doses (200–300 mg/kg) typically applied in rodent studies to test for reproductive effects. Some of these chemicals exert effects on rhesus monkey gonads, embryo or fetal development, and progeny phenotype, or in human reproductive tissues or stem cells.…”

Effects of Neonicotinoids on Promoter-Specific Expression and Activity of Aromatase: Implications for the Development of Hormone-Dependent Breast Cancer

“…Exposure to atrazine, a widely used herbicide, induces CYP19 expression, aromatase activity and estrogen biosynthesis in human cell lines [12,13,14,15,16] , but little is known about "emerging" contaminants such as neonicotinoid insecticides. Neonicotinoids are the most commonly used insecticides worldwide, and are applied as coatings to the seeds of corn, canola, soybeans and the majority of fruits and vegetables.…”