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

STAR Protoc. 2021 Apr 19;2(2):100463. doi: 10.1016/j.xpro.2021.100463. eCollection 2021 Jun 18.

Abstract

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).

Keywords: Cell Differentiation; Cell isolation; Neuroscience; Organoids; Stem Cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dopaminergic Neurons* / cytology
  • Dopaminergic Neurons* / metabolism
  • Humans
  • Mesencephalon* / cytology
  • Mesencephalon* / metabolism
  • Organoids* / cytology
  • Organoids* / metabolism
  • Pluripotent Stem Cells* / cytology
  • Pluripotent Stem Cells* / metabolism