MiR-21 regulates proliferation and invasion in trophoblastic cells and targets PTEN

Abstract

s / Placenta 34 (2013) A1–A99 A50 Objectives: Human trophoblast lineage-specific differentiation is difficult to study, due to a lack of a multipotent trophoblast stem cell (TSC) model. First trimester human cytotrophoblast (CTB) are bipotential, with the ability to differentiate into both hCG-secreting syncytiotrophoblast (STB) and HLAG-positive extravillous trophoblast (EVT); however, access to large numbers of such cells is limited. Human embryonic stem cells (hESCs) can be differentiated into trophoblast using BMP4 in presence of feederconditioned media (FCM), but these conditions are inconsistent and often result in heterogeneous populations of cells. We set out to develop defined media conditions for differentiation of hESCs into bipotential CTBs. Methods: Feeder-free hESCs (WA09/H9) were cultured in StemPro+bFGF. To differentiate the cells into CTB, cells were switched tominimalmedia for two days, then treated with BMP4 (10 ng/ml) for an additional four days. Microarray gene expression data from these cells were compared to data from first trimester primary CTB, placental stroma, amnion epithelial cells, JEG3 and BeWo cells, and human dermal fibroblasts. Treated hESCs were also evaluated by immunostaining for markers of pluripotency and CTB. Results: After four days in minimal media plus BMP4, cells downregulated the pluripotency marker OCT4, and induced CTB markers CK7, EGFR, and TP63. Microarray analysis revealed that these cells cluster most closely with JEG3 and BeWo and, in turn, are more closely related to the first trimester CTB. These cells could be replated, and when cultured in FCM+BMP4, could be induced to differentiate into both STB and EVT. Conclusions: In summary, we have successfully established a reproducible in vitro system, using defined media, to derive CTB-like cells from hESCs. These cells offer a superior alternative to the existing aneuploid human trophoblast cell lines, and, with the ability to differentiate into both STB and EVT, can be used to study trophoblast lineage-specific differentiation. http://dx.doi.org/10.1016/j.placenta.2013.06.151 P1.115. MIR-21 REGULATES PROLIFERATION AND INVASION IN TROPHOBLASTIC CELLS AND TARGETS PTEN Wittaya Chaiwangyen, Diana Morales-Prieto, Stephanie Ospina-Prieto, Ekkehard Schleussner, Udo Markert Placenta-Labor, Department of Obstetrics, University Hospital Jena, Jena,