Induction of human trophoblast stem-like cells from primed pluripotent stem cells

The placenta is a transient but important multifunctional organ crucial for healthy pregnancy for both mother and fetus. Nevertheless, limited access to human placenta samples and the paucity of a proper in vitro model system has hampered our understanding of the mechanisms underlying early human placental development and placenta-associated pregnancy complications. To overcome these constraints, we established a simple procedure with a short-term treatment of bone morphogenetic protein 4 (BMP4) in trophoblast stem cell culture medium (TSCM) to convert human primed pluripotent stem cells (PSCs) to trophoblast stem-like cells (TSLCs). These TSLCs show not only comparable morphology and gene expression profile to bona fide human trophoblast stem cells (TSCs) but also long-term self-renewal capacity with bipotency that allows the cells to differentiate into extravillous trophoblasts (EVT) and syncytiotrophoblasts (ST). These indicate that TSLCs are equivalent to genuine human TSCs. Our data suggest a straightforward approach to make human TSCs directly from pre-existing primed PSCs and provide a valuable opportunity to study human placenta development and pathology even from patients with placenta-related diseases. Overall design: RNA-seq was performed on the following cells: primary hTSCs (CT27), undifferentiated hESCs (H9), undifferentiated hiPSCs, hTSLCs (TS-like cells) derived from hESCs (ES-TSLCs), hTSLCs derived from hiPSCs (iPS-TSLCs), EVT differentiated from hTSCs, EVT differentiated from ES-TSLCs (ES-EVT), EVT differentiated from iPS-TSLCs (iPS-EVT), ST differentiated from hTSCs, ST differentiated from ES-TSLCs (ES-ST), and ST differentiated from iPS-TSLCs (iPS-ST). Two biological replicates were used for each cell type.