Protocol High-throughput generation of midbrain dopaminergic neuron organoids from reporter human pluripotent stem cells

Here, we describe a high-throughput 3D differentiation protocol for deriving midbrain dopaminergic neurons from human pluripotent stem cells. The use of organoids has become prevalent in disease modeling, but there is a high demand for more homogeneous cultures. Our approach is advantageous for large-scale production of uniform midbrain organoids that can be maintained in diverse formats, and our reporters allow for sorting of dopaminergic neurons. The maturing long-term organoid cultures can be used as a model for the entire midbrain. For complete details on the use and execution of this protocol, please refer to Ahfeldt et al. (2020).

Automated summary

This study presents a high-throughput 3D differentiation protocol for deriving midbrain dopaminergic neurons from human pluripotent stem cells, which allows for large-scale production of uniform midbrain organoids for disease modeling and dopaminergic neuron sorting. The long-term maturing organoid cultures can serve as a model for the entire midbrain.