2. Academic Validation
  3. Biphasic Activation of WNT Signaling Facilitates the Derivation of Midbrain Dopamine Neurons from hESCs for Translational Use

Biphasic Activation of WNT Signaling Facilitates the Derivation of Midbrain Dopamine Neurons from hESCs for Translational Use

  • Cell Stem Cell. 2021 Feb 4;28(2):343-355.e5. doi: 10.1016/j.stem.2021.01.005.
Tae Wan Kim 1,  Jinghua Piao 2,  So Yeon Koo 3,  Sonja Kriks 4,  Sun Young Chung 1,  Doron Betel 5,  Nicholas D Socci 6,  Se Joon Choi 7,  Susan Zabierowski 8,  Brittany N Dubose 8,  Ellen J Hill 8,  Eugene V Mosharov 7,  Stefan Irion 1,  Mark J Tomishima 8,  Viviane Tabar 9,  Lorenz Studer 10
Affiliations

Affiliations

  • 1 Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
  • 2 Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Department of Neurosurgery and Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
  • 3 Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Neuroscience Graduate Program of Weill Cornell Graduate School of Biomedical Sciences, Weill Cornell Medical College, New York, NY, USA.
  • 4 Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
  • 5 Institute for Computational Biomedicine, Division of Hematology/Oncology, Department of Medicine, Weill Cornell Medical College, New York, NY, USA.
  • 6 Bioinformatics Core, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
  • 7 Department of Neurology, Columbia University Medical Center, New York, NY, USA.
  • 8 Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA; SKI Stem Cell Research Facility, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
  • 9 Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Department of Neurosurgery and Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Cancer Biology & Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA. Electronic address: tabarv@mskcc.org.
  • 10 Center for Stem Cell Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA. Electronic address: studerl@mskcc.org.
PMID: 33545081 DOI: 10.1016/j.stem.2021.01.005
Abstract

Human pluripotent stem cells show considerable promise for applications in regenerative medicine, including the development of cell replacement paradigms for the treatment of Parkinson's disease. Protocols have been developed to generate authentic midbrain dopamine (mDA) neurons capable of reversing dopamine-related deficits in animal models of Parkinson's disease. However, the generation of mDA neurons at clinical scale suitable for human application remains an important challenge. Here, we present an mDA neuron derivation protocol based on a two-step Wnt signaling activation strategy that improves expression of midbrain markers, such as Engrailed-1 (EN1), while minimizing expression of contaminating posterior (hindbrain) and anterior (diencephalic) lineage markers. The resulting neurons exhibit molecular, biochemical, and electrophysiological properties of mDA neurons. Cryopreserved mDA neuron precursors can be successfully transplanted into 6-hydroxydopamine (6OHDA) lesioned rats to induce recovery of amphetamine-induced rotation behavior. The protocol presented here is the basis for clinical-grade mDA neuron production and preclinical safety and efficacy studies.

Keywords

Parkinson’s disease; WNT signaling; cell therapy; directed differentiation; human embryonic stem cells; human-induced pluripotent stem cells; midbrain development; neural patterning; preclinical study; transplantation.

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