title: Generation of a ciliary margin-like stem cell niche from self-organizing human retinal tissue.
abstract: In the developing neural retina (NR), multipotent stem cells within the ciliary margin (CM) contribute to de novo retinal tissue growth. We recently reported the ability of human embryonic stem cells (hESCs) to self-organize stratified NR using a three-dimensional culture technique. Here we report the emergence of CM-like stem cell niches within human retinal tissue. First, we developed a culture method for selective NR differentiation by timed BMP4 treatment. We then found that inhibiting GSK3 and FGFR induced the transition from NR tissue to retinal pigment epithelium (RPE), and that removing this inhibition facilitated the reversion of this RPE-like tissue back to the NR fate. This step-wise induction-reversal method generated tissue aggregates with RPE at the margin of central-peripherally polarized NR. We demonstrate that the NR-RPE boundary tissue further self-organizes a niche for CM stem cells that functions to expand the NR peripherally by de novo progenitor generation.
authors.affiliation: 1] Neurogenesis and Organogenesis Group, RIKEN Center for Developmental Biology, 2-2-3 Manatojima-Minamimachi, Chuo, Kobe 650-0047, Japan [2] Human Stem Cell Technology Unit, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo, Kobe 650-0047, Japan [3] Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., 3-1-98 Kasugade-naka, Konohana, Osaka 554-8558, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kuwahara
authors.affiliation: 1] Neurogenesis and Organogenesis Group, RIKEN Center for Developmental Biology, 2-2-3 Manatojima-Minamimachi, Chuo, Kobe 650-0047, Japan [2] Department of Endocrinology and Diabetes, Graduate School of Medicine, Nagoya University, 65 Tsurumai-cho, Showa, Nagoya 466-8550, Japan.
authors.forename: Chikafumi
authors.initials: C
authors.lastname: Ozone
authors.affiliation: 1] Neurogenesis and Organogenesis Group, RIKEN Center for Developmental Biology, 2-2-3 Manatojima-Minamimachi, Chuo, Kobe 650-0047, Japan [2] Human Stem Cell Technology Unit, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo, Kobe 650-0047, Japan [3] Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., 3-1-98 Kasugade-naka, Konohana, Osaka 554-8558, Japan.
authors.forename: Tokushige
authors.initials: T
authors.lastname: Nakano
authors.affiliation: Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., 3-1-98 Kasugade-naka, Konohana, Osaka 554-8558, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Saito
authors.affiliation: Four-Dimensional Tissue Analysis Unit, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo, Kobe 650-0047, Japan.
authors.forename: Mototsugu
authors.initials: M
authors.lastname: Eiraku
authors.affiliation: 1] Neurogenesis and Organogenesis Group, RIKEN Center for Developmental Biology, 2-2-3 Manatojima-Minamimachi, Chuo, Kobe 650-0047, Japan [2] Human Stem Cell Technology Unit, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo, Kobe 650-0047, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Sasai
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D005434:Flow Cytometry; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008813:Mice, Inbred ICR; D039902:Multipotent Stem Cells / Q000166:cytology; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000196:embryology*; D055213:Retinal Pigment Epithelium / Q000166:cytology; D055153:Stem Cell Niche / Q000502:physiology*
chemical_list: C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; C496348:FGFR1 protein, human; D051496:Receptor, Fibroblast Growth Factor, Type 1; D038362:Glycogen Synthase Kinase 3