A protocol for the differentiation of human embryonic stem cells into midbrain dopaminergic neurons

Here, we present a protocol for the generation of functional midbrain dopami-nergic (mDA) neurons from human embryonic stem cells (hESCs), which mimics the development of the human ventral midbrain. We describe steps for hESC pro-liferation, induction of mDA progenitors, freezing stocks of mDA progenitors as an intermediate starting point to reduce the time to make mDA neurons, and maturation of mDA neurons. The entire protocol is feeder-free and uses chemi-cally defined materials.For complete details on the use and execution of this protocol, please refer to Nishimura et al. (2023).1

Automated summary

This article introduces a method for generating functional midbrain dopaminergic (mDA) neurons from human embryonic stem cells (hESCs), which mimics the development of the human ventral midbrain. The protocol includes steps for hESC proliferation, induction of mDA progenitors, freezing stocks of mDA progenitors as an intermediate starting point to reduce the time to make mDA neurons, and maturation of mDA neurons. The entire method is feeder-free and utilizes chemically defined materials. For complete details on the use and execution of this protocol, please refer to Nishimura et al. (2023).