Transcriptional Dynamics of Cultured Human Villous Cytotrophoblasts.
During human pregnancy, cytotrophoblasts (CTBs) play key roles in uterine invasion, vascular remodeling, and anchoring of the feto-placental unit. Due to the challenges associated with studying human placentation in utero, cultured primary villous CTBs are used as a model of the differentiation pathway that leads to invasion of the uterine wall. In vitro, CTBs emulate in vivo cell behaviors, such as migration, aggregation, and substrate penetration. Although some of the molecular features related to these cell behaviors have been described, the underlying mechanisms, at a global level, remain undefined at midgestation. Thus, in this study, we characterized second-trimester CTB differentiation/invasion in vitro, correlating the major morphological transitions with the transcriptional changes that occurred at these steps. After plating on Matrigel as individual cells, CTBs migrated toward each other and formed multicellular aggregates. In parallel, using a microarray approach, we observed differentially expressed (DE) genes across time, which were enriched for numerous functions, including cell migration, vascular remodeling, morphogenesis, cell communication, and inflammatory signaling. DE genes encoded several molecules that we and others previously linked to critical CTB function in vivo, suggesting that the novel DE molecules we discovered played important roles. Immunolocalization confirmed that CTBs in situ gave a signal for two of the most highly expressed genes in vitro. In summary, we characterized, at a global level, the temporal dynamics of primary human CTB gene expression in culture. These data will enable future analyses of various types of in vitro perturbations-for example, modeling disease processes and environmental exposures.
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.|Eli and Edythe Broad Center for Regeneration Medicine and Stem Cell Research, University of California, San Francisco, California 94143.
Joshua F
JF
Robinson
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.|Eli and Edythe Broad Center for Regeneration Medicine and Stem Cell Research, University of California, San Francisco, California 94143.
Mirhan
M
Kapidzic
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.|Eli and Edythe Broad Center for Regeneration Medicine and Stem Cell Research, University of California, San Francisco, California 94143.
Matthew
M
Gormley
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.
Katherine
K
Ona
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.
Terrence
T
Dent
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.
Helia
H
Seifikar
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.
Emily G
EG
Hamilton
Center for Reproductive Sciences, University of California, San Francisco, California 94143.|Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, California 94143.|Eli and Edythe Broad Center for Regeneration Medicine and Stem Cell Research, University of California, San Francisco, California 94143.|Division of Maternal Fetal Medicine, University of California, San Francisco, California 94143.|Department of Anatomy, University of California, San Francisco, California 94143.|Human Embryonic Stem Cell Program, University of California, San Francisco, California 94143.
Susan J
SJ
Fisher
D002454:Cell Differentiation / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D010920:Placenta / Q000166:cytology / Q000378:metabolism; D010929:Placentation / Q000235:genetics; D011247:Pregnancy; D011261:Pregnancy Trimester, First / Q000235:genetics; D061251:Primary Cell Culture; D059467:Transcriptome; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism*