Generation of Functional Microglia from Human Pluripotent Stem Cells

Microglia are resident macrophages that are unique in the central nervous system (CNS), including the brain, spinal cord, and retina. Microglia play a critical role in the development and homeostasis of the CNS. Therefore, microglia have become a focus of research on the pathogenesis of neurological diseases. Due to the limited source of primary human microglia, previous studies of microglia function have been conducted mainly in animal models. However, there are significant differences between mouse and human microglia, especially the lack of homologous genes. In this chapter, we provide a detailed, step-by-step, easy-to-follow protocol to obtain highly pure (more than 90%) human microglia in around 60 days from human embryonic stem cells and human induced pluripotent stem cells without sorting, using three cytokines (M-CSF, IL-3, and IL-34). It is hoped that this protocol will be beneficial for disease modeling and regenerative medicine, and will contribute to in-depth studies of the pathogenesis and therapeutics of CNS-related diseases.