Differentiation of Human Induced Pluripotent Stem Cells (hiPSC) into Endothelial-Type Cells and Establishment of an...

Development of central nervous system (CNS) therapeutics and their brain delivery is impeded by the presence of the blood-brain barrier (BBB). In vitro BBB models, in particular human in vitro BBB models, are critical tools for CNS drug research and development. However, the availability of primary human microvascular endothelial cells is very limited for in vitro modeling. Advances in human induced pluripotent stem cell (hiPSC) technologies provide reproducible human cell resources for scientific research, regenerative medicine, and in vitro modeling. In particular, the differentiation of hiPSC into brain endothelial cells provides scalable, renewable and unlimited cells for in vitro BBB modeling that enables rapid screening of CNS drugs in terms of their BBB permeability. The following protocols provide a general guideline for hiPSC culture, differentiation of hiPSC into endothelial cells (hiPSC-ECs), generation of rat primary astrocytes, and establishment of a two-chamber co-culture in vitro BBB model.