title: Efficient generation of region-specific forebrain neurons from human pluripotent stem cells under highly defined condition.
abstract: Human pluripotent stem cells (hPSCs) have potential to differentiate to unlimited number of neural cells, which provide powerful tools for neural regeneration. To date, most reported protocols were established with an animal feeder system. However, cells derived on this system are inappropriate for the translation to clinical applications because of the introduction of xenogenetic factors. In this study, we provided an optimized paradigm to generate region-specific forebrain neurons from hPSCs under a defined system. We assessed five conditions and found that a vitronectin-coated substrate was the most efficient method to differentiate hPSCs to neurons and astrocytes. More importantly, by applying different doses of purmorphamine, a small-molecule agonist of sonic hedgehog signaling, hPSCs were differentiated to different region-specific forebrain neuron subtypes, including glutamatergic neurons, striatal medium spiny neurons, and GABA interneurons. Our study offers a highly defined system without exogenetic factors to produce human neurons and astrocytes for translational medical studies, including cell therapy and stem cell-based drug discovery.
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Yuan
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Kai-Heng
authors.initials: KH
authors.lastname: Fang
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Shi-Ying
authors.initials: SY
authors.lastname: Cao
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Zhuang-Yin
authors.initials: ZY
authors.lastname: Qu
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Ophthalmology, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Krencik
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Min
authors.initials: M
authors.lastname: Xu
authors.affiliation: Waisman Center, University of Wisconsin, Madison, WI, USA.
authors.forename: Anita
authors.initials: A
authors.lastname: Bhattacharyya
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Yu-Wen
authors.initials: YW
authors.lastname: Su
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Dong-Ya
authors.initials: DY
authors.lastname: Zhu
authors.affiliation: Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D006801:Humans; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D016548:Prosencephalon / Q000166:cytology* / Q000378:metabolism