title: Hippo/YAP-mediated rigidity-dependent motor neuron differentiation of human pluripotent stem cells.
abstract: Our understanding of the intrinsic mechanosensitive properties of human pluripotent stem cells (hPSCs), in particular the effects that the physical microenvironment has on their differentiation, remains elusive. Here, we show that neural induction and caudalization of hPSCs can be accelerated by using a synthetic microengineered substrate system consisting of poly(dimethylsiloxane) micropost arrays (PMAs) with tunable mechanical rigidities. The purity and yield of functional motor neurons derived from hPSCs within 23 days of culture using soft PMAs were improved more than fourfold and tenfold, respectively, compared with coverslips or rigid PMAs. Mechanistic studies revealed a multi-targeted mechanotransductive process involving Smad phosphorylation and nucleocytoplasmic shuttling, regulated by rigidity-dependent Hippo/YAP activities and actomyosin cytoskeleton integrity and contractility. Our findings suggest that substrate rigidity is an important biophysical cue influencing neural induction and subtype specification, and that microengineered substrates can thus serve as a promising platform for large-scale culture of hPSCs.
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Yubing
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Koh Meng Aw
authors.initials: KM
authors.lastname: Yong
authors.affiliation: 1] Department of Biologic and Materials Sciences, University of Michigan, Ann Arbor, Michigan 48109, USA [2] Biointerfaces Institute, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Luis G
authors.initials: LG
authors.lastname: Villa-Diaz
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Weiqiang
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Renee
authors.initials: R
authors.lastname: Philson
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Shinuo
authors.initials: S
authors.lastname: Weng
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Haoxing
authors.initials: H
authors.lastname: Xu
authors.affiliation: 1] Department of Biologic and Materials Sciences, University of Michigan, Ann Arbor, Michigan 48109, USA [2] Biointerfaces Institute, University of Michigan, Ann Arbor, Michigan 48109, USA [3] Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Paul H
authors.initials: PH
authors.lastname: Krebsbach
authors.affiliation: 1] Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA [2] Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Jianping
authors.initials: J
authors.lastname: Fu
mesh_terms: D000205:Actomyosin / Q000378:metabolism; D018797:Cell Cycle Proteins; D002454:Cell Differentiation; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D000090683:Hippo Signaling Pathway; D006801:Humans; D040542:Mechanotransduction, Cellular; D009046:Motor Neurons / Q000166:cytology / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051785:Smad Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D051785:Smad Proteins; D014157:Transcription Factors; C456659:YY1AP1 protein, human; D000205:Actomyosin; D017346:Protein Serine-Threonine Kinases
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