Orchestrated trophoblast differentiation is necessary to establish and maintain a normal pregnancy, however the molecular mechanisms that guide this process remain largely unknown. Although early studies of cytotrophoblast differentiation relied on animal models, more recent trophoblast research has involved in vitro models of human tissue. These in vitro models have utilized cultured trophoblast cell lines, primary cell culture, and villous explant cultures-each with its advantages and disadvantages. Traditionally, attempts to develop in vitro models of human placental differentiation have relied on two-dimensional cell culture. Though monolayer culture methods have been refined over time this technique has several limitations, including the inability to study cell-to-cell interactions. Recently, several studies have employed three-dimensional culture methods to overcome many of the limitations of traditional two-dimensional trophoblast culture. These three-dimensional culture systems have an important role in both the study of cytotrophoblast differentiation and development of new therapeutics targeting placenta associated diseases.
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