Cota-Coronado et al. (2020). STAR Protocols. doi: 10.1016/j.xpro.2020.100065
…Cruz Biotechnology Sc-990, RRID:AB_2298676 Goat polyclonal anti-OTX2 R&D Systems AF1979, RRID:AB_2157172 Sheep polyclonal anti-PITX2 R&D Systems AF7388, RRID:AB_11128639 Rabbit polyclonal anti-TH Pel-Freez Biologicals P40101-0, RRID:AB_461064 Sheep polyclonal anti-TH Pel-Freez Biologicals P60101-0, RRID:AB_461070 Chemicals, Peptides, and Recombinant Proteins Accutase Innovative Cell Technologies AT104 Ascorbic…
Pel-Freez products cited: Rabbit Anti-Tyrosine Hydroxylase (P40101), Sheep Anti-Tyrosine Hydroxylase (P60101)
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) and Niclis et al. (2017a).