Optimized Culture and Decidualization of Trophoblast Cells

LAD2 cells (kindly provided by Dr. Kirschbaum, passages 3 - 10) were cultured in StemPro-34 containing recombinant human stem cell factor (SCF; Peprotech, Rocky Hill, NJ, USA) (25 (link)). HTR-8/SVneo cells (kindly provided by Dr. Graham, passage 3 - 12) (26 (link)) and T HESCs (purchased from the American Type Culture Collection; Manassas, VA, USA, passages 3 - 10) were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin. Decidualization of T HESCs was induced by culturing with 1 µM medroxyprogesterone acetate (MPA; #M1629; Sigma-Aldrich, St. Louis, MO, USA) and 0.5 mM 8-bromoadenosine 3’:5’-cyclic monophosphate (8-Br cAMP; #B7880; Sigma) for 5–6 days (27 (link)). We confirmed the decidualization of T HESCs based on the presence of morphological changes and increased PRL mRNA.