title: SOX10-Nano-Lantern Reporter Human iPS Cells; A Versatile Tool for Neural Crest Research.
abstract: The neural crest is a source to produce multipotent neural crest stem cells that have a potential to differentiate into diverse cell types. The transcription factor SOX10 is expressed through early neural crest progenitors and stem cells in vertebrates. Here we report the generation of SOX10-Nano-lantern (NL) reporter human induced pluripotent stem cells (hiPS) by using CRISPR/Cas9 systems, that are beneficial to investigate the generation and maintenance of neural crest progenitor cells. SOX10-NL positive cells are produced transiently from hiPS cells by treatment with TGFβ inhibitor SB431542 and GSK3 inhibitor CHIR99021. We found that all SOX10-NL-positive cells expressed an early neural crest marker NGFR, however SOX10-NL-positive cells purified from differentiated hiPS cells progressively attenuate their NL-expression under proliferation. We therefore attempted to maintain SOX10-NL-positive cells with additional signaling on the plane and sphere culture conditions. These SOX10-NL cells provide us to investigate mass culture with neural crest cells for stem cell research.
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Horikiri
authors.affiliation: Department of Biomedical Engineering, Faculty of Life Sciences, Doshisha University, Kyotanabe, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ohi
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
authors.forename: Mitsuaki
authors.initials: M
authors.lastname: Shibata
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Ikeya
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Morio
authors.initials: M
authors.lastname: Ueno
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Chie
authors.initials: C
authors.lastname: Sotozono
authors.affiliation: Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kinoshita
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.|Advanced Biomedical Engineering Research Center, Doshisha University, Kyotanabe, Japan.
authors.forename: Takahiko
authors.identifier: http://orcid.org/0000-0003-3836-7978
authors.initials: T
authors.lastname: Sato
mesh_terms: D002454:Cell Differentiation; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D017930:Genes, Reporter; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism*; D055757:SOXE Transcription Factors / Q000235:genetics*
chemical_list: D055757:SOXE Transcription Factors