title: Simple Derivation of Spinal Motor Neurons from ESCs/iPSCs Using Sendai Virus Vectors.
abstract: Amyotrophic lateral sclerosis (ALS) is a progressive and fatal degenerative disorder of motor neurons (MNs). Embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs) now help us to understand the pathomechanisms of ALS via disease modeling. Various methods to differentiate ESCs/iPSCs into MNs by the addition of signaling molecules have been reported. However, classical methods require multiple steps, and newer simple methods using the transduction of transcription factors run the risk of genomic integration of the vector genes. Heterogeneity of the expression levels of the transcription factors also remains an issue. Here we describe a novel approach for differentiating human and mouse ESCs/iPSCs into MNs using a single Sendai virus vector encoding three transcription factors, LIM/homeobox protein 3, neurogenin 2, and islet-1, which are integration free. This single-vector method, generating HB9-positive cells on day 2 from human iPSCs, increases the ratio of MNs to neurons compared to the use of three separate Sendai virus vectors. In addition, the MNs derived via this method from iPSCs of ALS patients and model mice display disease phenotypes. This simple approach significantly reduces the efforts required to generate MNs, and it provides a useful tool for disease modeling.
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, Kyoto 6068507, Japan; Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 6068507, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Goto
authors.affiliation: Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 6068507, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Imamura
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, Kyoto 6068507, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Komatsu
authors.affiliation: Division of Gene Therapy and Genome Editing, Research Center for Genomic Medicine, Saitama Medical University, Saitama 3501241, Japan.
authors.forename: Kohnosuke
authors.initials: K
authors.lastname: Mitani
authors.affiliation: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 6068501, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Aiba
authors.affiliation: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 6068501, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Nakatsuji
authors.affiliation: DNAVEC Center, ID Pharma Co., Ltd., Tsukuba 3002611, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Inoue
authors.affiliation: Department of Neurology, Tokyo Metropolitan Neurological Hospital, Tokyo 1830042, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kawata
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, Kyoto 6068507, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Yamashita
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, Kyoto 6068507, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Takahashi
authors.affiliation: Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 6068507, Japan.
authors.forename: Haruhisa
authors.initials: H
authors.lastname: Inoue
keywords: ESC; Sendai virus; differentiation; direct conversion; embryonic stem cells; iPSC; induced pluripotent stem cells; motor neurons; transcription factor