Co-culture of JEG-3, BeWo and syncBeWo cell lines with adrenal H295R cell line: an alternative model for examining endocrine and metabolic properties of the fetoplacental unit

Abstract

Monocultures of different placental cells are used for many physiological and toxicological studies; however, they are not a true reflection of the interaction between placenta and fetus. To develop the most appropriate model to study endocrine and metabolic properties of fetoplacental unit we used three co-culture models of placental cells nonfusogenic JEG-3, unsyncytialised BeWo (BeWo) and syncytialised BeWo (syncBeWo) cultured with adrenal (H295R) cells. As an end point of endocrine properties we investigated steroids receptors expression and steroid secretion, while as metabolic properties AhR, CYP1A1and COMT expression. Progesterone (P4), estradiol (E2) and human chorionic gonadotropin (hCG) secretion (ELISA) and 3βHSD, CYP19, estrogen (ERα/β), progesterone (PR) and aryl hydrocarbon (AhR) receptors, CYP1A1 and COMT protein expression (Western blot) were evaluated. Comparing three co-culture models we observed: (1) there were no differences between JEG-3 and BeWo in the PR expression, however it was higher in BeWo compared to syncBeWo; (2) there were no differences in ERα protein expression in all models, while profile of ERβ expression was the highest in syncBeWo; (3) high P4 secretion in JEG-3 and BeWo while low in syncBeWo; (4) high E2 levels in JEG-3 and syncBeWo, while low E2 secretion in BeWo; (5) the highest hCG secretion in the JEG-3 and syncBeWo than in BeWo (6) the highest AhR, CYP1A1 and COMT expression in syncBeWo. Based on the results showing higher hCG secretion in the JEG-3 than in BeWo, representing villous and extravillous phenotype we suggest that JEG-3 model could be used to study fetoplacental steroidogenesis at the 1st, while BeWo model at the 3rd. Results showing comparable profiles of AhR, CYP1A1 and COMT expression in JEG-3 and BeWo models and the significantly higher expression in synBeWo points to synBeWo as a good model for study the metabolic properties.Electronic supplementary materialThe online version of this article (doi:10.1007/s10616-017-0142-z) contains supplementary material, which is available to authorized users.

Toxicity of anticancer drugs in human placental tissue explants and trophoblast cell lines

“…Additionally, abnormal steroidogenesis has been associated with placenta-related adverse pregnancy outcomes, such as preeclampsia (Berkane et al 2018 ). Here, we studied inhibition of steroidogenesis in syncytialised BeWo cells successfully and found that the increase in progesterone release after forskolin treatment is in line with previous studies (Drwal et al 2018 ). From a methodological point of view, we added to this that the amount of DMSO interferes with forskolin-induced progesterone release.…”

“…7) after 48 h exposure. Statistical analysis was performed with a one-way ANOVA and means were compared to control using the Bonferroni post-hoc test *P < 0.05, ***P < 0.001 treatment is in line with previous studies (Drwal et al 2018). From a methodological point of view, we added to this that the amount of DMSO interferes with forskolin-induced progesterone release.…”

“…Although the placenta seems to be capable of de novo estrogen synthesis from cholesterol present in FCS (Escobar et al 2011 ), estrogen production is generally thought to be largely dependent on fetal androgens. Addition of precursor androgens achieved by co-culturing adrenal cells with syncytialised BeWo cells was shown to increase estrogen synthesis substantially (Drwal et al 2018 ). In term placental explants, a decrease was found of progesterone release of approximately 50-fold over the first four days of culture (Sato et al 2015 ).…”

Physiology and Pathophysiology of Steroid Biosynthesis, Transport and Metabolism in the Human Placenta

“…In this context the recently established co-culture models of choriocarcinoma cells (JEG-3 or BeWo) with H295R cells should be considered for future analysis. The H295R/BeWo co-culture model offers the opportunity to evaluate the effects of chemical exposures on androgen and estrogen biosynthesis, as well as on various other aspects of feto–placental communication ( Hudon Thibeault et al, 2017 ; Drwal et al, 2018 ; Thibeault et al, 2018 ).…”

Culture strategy as a modulator of target assessments: Functionality of suspension versus hanging drop‐derived choriocarcinoma spheroids as in vitro model of embryo implantation

“…Choriocarcinoma cell lines (BeWo and JEG‐3 cell lines), as malignant counterparts of trophoblast, have characteristic similarities to the trophoblastic part of an embryo 22 . Apart from endocrine function, proliferation, invasive, attachment, and adhesion potentials of two cell lines, approximately 2700 genes are differentially expressed between them 1,3,22 . Various characteristics of heterogeneous BeWo and JEG‐3 cell lines make them identical to specific experimental treatment 3,10,23 .…”