Robinson Joshua F, Kapidzic Mirhan, Gormley Matthew, Ona Katherine, Dent Terrence, Seifikar Helia, Hamilton Emily G, Fisher Susan J
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.
Endocrinology. 2017 Jun 1;158(6):1581-1594. doi: 10.1210/en.2016-1635.
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.
在人类妊娠期间，细胞滋养层细胞（CTBs）在子宫侵袭、血管重塑以及胎儿 - 胎盘单位的锚定过程中发挥着关键作用。由于在子宫内研究人类胎盘形成存在诸多挑战，培养的原代绒毛CTBs被用作研究导致子宫壁侵袭的分化途径的模型。在体外，CTBs能够模拟体内细胞行为，如迁移、聚集和穿透基质。尽管已经描述了一些与这些细胞行为相关的分子特征，但在妊娠中期，从整体水平来看，其潜在机制仍不明确。因此，在本研究中，我们对孕中期CTBs的体外分化/侵袭进行了表征，将主要的形态学转变与这些步骤中发生的转录变化相关联。在以单个细胞接种到基质胶上后，CTBs相互迁移并形成多细胞聚集体。同时，我们采用微阵列方法观察到不同时间点差异表达（DE）的基因，这些基因在众多功能中富集，包括细胞迁移、血管重塑、形态发生、细胞通讯和炎症信号传导。DE基因编码了一些我们和其他人之前在体内与关键CTB功能相关联的分子，这表明我们发现的新DE分子发挥了重要作用。免疫定位证实原位CTBs对体外两个表达最高的基因发出信号。总之，我们在整体水平上表征了培养的原代人CTB基因表达的时间动态。这些数据将有助于未来对各种类型体外干扰的分析，例如模拟疾病过程和环境暴露。
