In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

作者：Wang Jianle; Anguera Montserrat C^*

来源：Jove-Journal of Visualized Experiments, 2017, (121): e55268.

DOI：10.3791/55268

• Developmental Biology
• Issue 121
• human trophoblastic cells
• pluripotent stem cells
• human embryonic stem cells
• human induced pluripotent stem cells
• in vitro differentiation
• bone morphogenic protein 4
• Activin/Nodal pathways

The placenta is the first organ to develop during embryogenesis and is required for the survival of the developing embryo. The placenta is comprised of various trophoblastic cells that differentiate from the extra-embryonic trophectoderm cells of the preimplantation blastocyst. As such, our understanding of the early differentiation events of the human placenta is limited because of ethical and legal restrictions on the isolation and manipulation of human embryogenesis. Human pluripotent stem cells (hPSCs) are a robust model system for investigating human development and can also be differentiated in vitro into trophoblastic cells that express markers of the various trophoblast cell types. Here, we present a detailed protocol for differentiating hPSCs into trophoblastic cells using bone morphogenic protein 4 and inhibitors of the Activin/Nodal signaling pathways. This protocol generates various trophoblast cell types that can be transfected with siRNAs for investigating loss-of-function phenotypes or can be infected with pathogens. Additionally, hPSCs can be genetically modified and then differentiated into trophoblast progenitors for gain-of-function analyses. This in vitro differentiation method for generating human trophoblasts starting from hPSCs overcomes the ethical and legal restrictions of working with early human embryos, and this system can be used for a variety of applications, including drug discovery and stem cell research.

• 出版日期2017-3