Journal article
An Optimized Protocol for the Generation of Midbrain Dopamine Neurons under Defined Conditions
CW Gantner, A Cota-Coronado, LH Thompson, CL Parish
STAR Protocols | Published : 2020
Abstract
Here, we describe a xeno-free, feeder-free, and chemically defined protocol for the generation of ventral midbrain dopaminergic (vmDA) progenitors from human pluripotent stem cells (hPSCs). This simple-to-follow protocol results in high yields of cryopreservable dopamine neurons across multiple hPSC lines. Wnt signaling is the critical component of the differentiation and can be finely adjusted in a line-dependent manner to enhance production of dopamine neurons for the purposes of transplantation, studying development and homeostasis, disease modeling, drug discovery, and drug development. For complete details on the use and execution of this protocol, please refer to Gantner et al. (2020) ..
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Awarded by State Government of Victoria
Funding Acknowledgements
The authors thank current and past members of the Thompson and Parish laboratories for contin-uous discussion and refinement of the protocol. In particular, we thank Jonathan Niclis, Isabelle de Luzy, and Jennifer Durnall for their contribution. C.W.G. was supported by an Australian Post-graduate Award. C.L.P. was supported by a National Health and Medical Research Council (NHMRC) Australia, Senior Research Fellowship. This work was supported by an NHMRC Australia project grant (APP1102704 to C.L.P.) and Stem Cells Australia (to C.L.P.) . The Florey Institute of Neurosci-ence and Mental Health acknowledges the strong support from the Victorian Government and in particular the funding from the Operational Infrastructure Support Grant.