[PubMed] [CrossRef] [Google Scholar] 39. in TCL. These results establish CCK, gastrin, and their receptor(s) in both human and mouse placentas, and TAS2R14 in the human placenta. Both CCK and TAS2R14 agonists increased intracellular calcium in human TCL. Although the functions of these ligands and receptors, and their potential cross talk in normal and pathological placentas, are currently unknown, this study opens new avenues for placental research. expression relative to normal pregnancy [“type”:”entrez-geo”,”attrs”:”text”:”GSE40182″,”term_id”:”40182″GSE40182 (52) and (32)]. In fact, as portrayed in Fig. 1 (previously unpublished), by using bioinformatics approaches similar to those we earlier described (28), was found to be one of the few differentially expressed genes in common between first-trimester CVS and delivered placentas from women afflicted with preeclampsia. Therefore, we reasoned that this virtually unique confluence could be more than coincidental, and placental CCK may be of particular biological significance and worthy of further concern. CCK mRNA was previously identified in placentas of women and mice during genome-wide gene expression profiling and showed increased expression in cells of unidentified origin in the metrial region of placentas from diabetic mice by in situ hybridization (37, 39). Otherwise, to the best of our knowledge, placental CCK has received little, if any, investigative attention. Open in a separate windows Fig. 1. Overlap of differentially expressed genes (DEG) between preeclamptic (PE) and normal pregnancies in chorionic villous samples (CVS) and delivered placentas. and and in our CVS microarray (“type”:”entrez-geo”,”attrs”:”text”:”GSE12767″,”term_id”:”12767″GSE12767 but not of other genes in the microarray: or and CmRNA in mouse placenta. Although a complete understanding of the physiological and potential pathophysiological functions of CCK, gastrin, and TAS2R14, as well as their potential interactions in the placenta require further investigation, herein we establish CCK, GAST, and CCK receptor expression by human and mouse trophoblasts, TAS2R14 expression by human trophoblasts, and further demonstrate that CCK and TASR14 receptor ligands are both coupled to calcium signaling, CCK mostly, if not exclusively, through CCKBR. MIS MATERIALS AND METHODS Human placentas. Human placentas were obtained from uncomplicated pregnancies after AMG-Tie2-1 delivery under Institutional Review Board (IRB) approval from the University of Florida (IRB 201602330). Tissue cryosections of first- and second-trimester placentas were provided by the National Institutes of Health Placental Bank at the University of California San Francisco (NICHD/NIH HD055764) under IRB approval from the University of Florida (IRB 201600936). Human trophoblast cell lines. Trophoblast cell lines were propagated in T75 flasks at 37C under standard tissue culture conditions (5% CO2-balance room air). The HTR-8/SVneo trophoblast cell line derived from first-trimester human villous trophoblasts was generously provided by Dr. Charles Graham, Queens University (12). We previously corroborated the trophoblast origin of these cells in our laboratory (26). HTR-8/SVneo cells were cultured in T75 flasks made up of complete media: RPMI 1640 with glucose (2.0 g/l), l-glutamine (0.3 g/l), and sodium bicarbonate (2.0 AMG-Tie2-1 g/l). JAR, JEG, and BeWo choriocarcinoma cells are trophoblast derived and they were purchased from the American Type Culture Collection. JAR cells were cultured in the AMG-Tie2-1 same media as described for the HTR-8/SVneo cells. JEG cells were cultured in EMEM made up of AMG-Tie2-1 l-glutamine (0.3 g/l) and sodium pyruvate (0.1 g/l). BeWo cells were propagated in F12 Nutrient Mixture Kaighns Modification with l-glutamine (0.3 g/l). All media were supplemented with penicillin (100 U/ml), streptomycin (100 g/ml; Sigma, St. Louis, MO, or.