Biphasic Activation of WNT Signaling Facilitates the Derivation of Midbrain Dopamine Neurons from hESCs for Translational Use v1

• DOI: 10.17504/protocols.io.bu7znzp6
• Publication Date: 2021-08-19T18:08:58Z
• Authors: Tae Wan Kim; Jinghua Piao; So Yeon Koo; Sonja Kriks; Sun Young Chung; Doron Betel; Nicholas D. Socci; Se Joon Choi; Susan Zabierowski; Brittany N. Dubose; Ellen J. Hill; Eugene V. Mosharov; Stefan Irion; Mark J. Tomishima; Viviane Tabar; Lorenz Studer
• Keywords: Hindbrain

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