Modeling Aspects of Human Placental Development with Trophoblast Stem Cells.

Human trophoblast stem (TS) cells provide a powerful in vitro model to study trophoblast lineage specification, placental development, and pregnancy-related disorders. This chapter presents detailed methodologies for the maintenance, derivation, and differentiation of TS cells, with a focus on reproducibility. We describe optimized culture conditions that preserve stem cell identity, including culture medium composition, extracellular matrix requirements, and passaging techniques. Step-by-step protocols for inducing differentiation into extravillous trophoblast and syncytiotrophoblast are presented, along with morphological, molecular, and functional assays to assess lineage commitment. Additionally, we describe a procedure to generate organoids from TS cells. Practical guidance is provided for maintaining reproducibility across experiments. These protocols can be implemented to investigate the role of diverse proteins, including members of the ADAMTS family, in human placental development. By enabling reproducible and controlled TS cell expansion and trophoblast cell differentiation, this methodology supports both basic research and disease modeling.