title: Transplantation of neurons derived from human iPS cells cultured on collagen matrix into guinea-pig cochleae.
abstract: The present study examined the efficacy of a neural induction method for human induced pluripotent stem (iPS) cells to eliminate undifferentiated cells and to determine the feasibility of transplanting neurally induced cells into guinea-pig cochleae for replacement of spiral ganglion neurons (SGNs). A stepwise method for differentiation of human iPS cells into neurons was used. First, a neural induction method was established on Matrigel-coated plates; characteristics of cell populations at each differentiation step were assessed. Second, neural stem cells were differentiated into neurons on a three-dimensional (3D) collagen matrix, using the same protocol of culture on Matrigel-coated plates; neuron subtypes in differentiated cells on a 3D collagen matrix were examined. Then, human iPS cell-derived neurons cultured on a 3D collagen matrix were transplanted into intact guinea-pig cochleae, followed by histological analysis. In vitro analyses revealed successful induction of neural stem cells from human iPS cells, with no retention of undifferentiated cells expressing OCT3/4. After the neural differentiation of neural stem cells, approximately 70% of cells expressed a neuronal marker, 90% of which were positive for vesicular glutamate transporter 1 (VGLUT1). The expression pattern of neuron subtypes in differentiated cells on a 3D collagen matrix was identical to that of the differentiated cells on Matrigel-coated plates. In addition, the survival of transplant-derived neurons was achieved when inflammatory responses were appropriately controlled. Our preparation method for human iPS cell-derived neurons efficiently eliminated undifferentiated cells and contributed to the settlement of transplant-derived neurons expressing VGLUT1 in guinea-pig cochleae. Copyright © 2015 John Wiley & Sons, Ltd.
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Ishikawa
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Hiroe
authors.initials: H
authors.lastname: Ohnishi
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Desislava
authors.initials: D
authors.lastname: Skerleva
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Tatsunori
authors.initials: T
authors.lastname: Sakamoto
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Yamamoto
authors.affiliation: Centre for iPS Cell Research and Application, Kyoto University, Japan.
authors.forename: Akitsu
authors.initials: A
authors.lastname: Hotta
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Juichi
authors.initials: J
authors.lastname: Ito
authors.affiliation: Department of Otolaryngology, Head and Neck Surgery, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Nakagawa
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D018914:Cells, Immobilized / Q000166:cytology / Q000637:transplantation; D003051:Cochlea; D003094:Collagen / Q000737:chemistry*; D005109:Extracellular Matrix / Q000737:chemistry*; D006168:Guinea Pigs; D064593:Heterografts; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000637:transplantation
chemical_list: D003094:Collagen
keywords: glutamatergic neuron; hearing; inner ear; neural induction; pluripotent stem cell; regeneration; scaffold; spiral ganglion neuron