JEG-3 placental cells in toxicology studies: a promising tool to reveal pregnancy disorders

Abstract

Placental alterations are responsible for adverse pregnancy outcomes like preeclampsia and intrauterine growth restriction. And yet, placenta toxicology has not become a fully-fledged toxicology field. Because placenta is very often seen only as a barrier between the mother and the fetus, there is a lack and therefore a need for an experimental human model with technical recommendations to study placenta toxicology. In vitro approaches are recommended in experimental toxicology as they focus on a specific biological process and yield high-throughput screening methods. In the present study, we first established incubation conditions to preserve signatures of the human JEG-3 cell line identity while enabling toxicity detection. JEG-3 cells prepared in our incubation conditions were renamed JEG-Tox cells. As placental alterations are mainly triggered by uncontrolled apoptosis, we second used known apoptotic agents pregnant women are exposed to, to check that JEG-Tox cells can trigger apoptosis. Ethanol, bisphenol F, quinalphos, 4,4’-DDT, benzalkonium chloride, phenoxyethanol, propylparaben, and perfluorooctanic acid all induced chromatin condensation in JEG-Tox cells. Our incubation conditions allow JEG-Tox cells to keep placental cell identity and to respond to toxic chemicals. JEG-Tox cells are a pertinent model for placenta toxicology and could be used to better understand pregnancy alterations.

Pregnant Women and Endocrine Disruptors: Role of P2X7 Receptor and Mitochondrial Alterations in Placental Cell Disorders

“…Cell incubation: To study the P2X7 receptor and its relationship between apoptosis, the cells were preincubated with either PBS or P2X7 antagonist Brilliant Blue G (BBG) at 25 µM for 15 min [34]. After removal of PBS and BBG, the cells were incubated for 72 h with bisphenol A (5, 10 and 20 µM), diethylstilbestrol (3.75, 7.5 and 15 µM), 4-tert-amylphenol (1, 10 and 50 µM), 4-heptylphenol (1, 10 and 50 µM), triclosan (0.1, 1 and 10 µM), propylparaben (20, 50 and 100 µM), benzyl butyl phthalate (1, 10 and 50 µM), DEHP (1, 10 and 50 µM) and 3-benzylidene camphor (1, 10 and 50 µM) in MEM with 2.5% FBS according to Olivier et al's protocol that describes the JEG-Tox model [35] or DMEM with 2.5% FBS for the HaCaT and A549. Concentrations tested in placental cells were selected according to the literature and the same concentrations were used to study the lung and skin cells [6,7,9,32,36,37].…”

Regulatory Testing for Endocrine Disruptors; Need for Validated Methods and Integrated Approaches

“…For example, the first selected method (Rat et al, 2017;Olivier et al, 2021), based on the use of human placental cells for the measurement of P2X7 activation, estradiol, progesterone, hPlacental Lactogen, and hyperglycosylated βhCG secretions, could be described as.…”

“…For example, the first selected method ( Rat et al, 2017 ; Olivier et al, 2021 ), based on the use of human placental cells for the measurement of P2X7 activation, estradiol, progesterone, hPlacental Lactogen, and hyperglycosylated βhCG secretions, could be described as. • having a TRC of 75% of the maximum, • being on level 2 of the OECD Conceptual Framework, • addressing early/intermediate Key Events • addressing a knowledge gap on female reproduction/fertility via placental function …”

Identification of Adrenomedullin-Induced S-Nitrosylated Proteins in JEG-3 Placental Cells

“…Cell Culture and Cell Lines JEG-3 is a choriocarcinoma cell line (HTB-36™, ATCC, USA) derived from human placenta. It shows EVCT characteristics including expression of human leukocyte antigen G, and is frequently employed as an EVCT model in studies on trophoblast invasion and migration in vitro [8,9]. The cells were authenticated and tested for contamination.…”