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.
Highlights
Based on the results showing higher human chorionic gonadotropin (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
Cell-lines derived from human placenta and chorion, such as human choriocarcinoma cells BeWo or JEG-3 are an alternative for study the placenta function (Sullivan 2002)
These differences in gene expression patterns suggest that JEG3 and BeWo cell lines will vary in their capacity to respond to an identical experimental treatment (Burleigh et al 2007)
Summary
Cell-lines derived from human placenta and chorion, such as human choriocarcinoma cells BeWo or JEG-3 are an alternative for study the placenta function (Sullivan 2002). Microarray analysis has indicated that approximately 2700 genes are differentially expressed between BeWo and JEG-3 cells, suggesting that they are suited only for specific experimental paradigms. These differences in gene expression patterns suggest that JEG3 and BeWo cell lines will vary in their capacity to respond to an identical experimental treatment (Burleigh et al 2007). Few papers indicate that significant differences in gene expression promotes fusion such as: b subunit of human chorionic gonadotropin, placental alkaline phosphatase, LGALS13, syncytin-1 or syncytin-2 in BeWo cells treatment with forskolin compared to non-treated BeWo cells (Orendi et al 2010; Frendo et al 2003)
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