title: Generation of Integration-free Induced Neural Stem Cells from Mouse Fibroblasts.
abstract: The viral vector-mediated overexpression of the defined transcription factors, Brn4/Pou3f4, Sox2, Klf4, and c-Myc (BSKM), could induce the direct conversion of somatic fibroblasts into induced neural stem cells (iNSCs). However, viral vectors may be randomly integrated into the host genome thereby increasing the risk for undesired genotoxicity, mutagenesis, and tumor formation. Here we describe the generation of integration-free iNSCs from mouse fibroblasts by non-viral episomal vectors containing BSKM. The episomal vector-derived iNSCs (e-iNSCs) closely resemble control NSCs, and iNSCs generated by retrovirus (r-iNSCs) in morphology, gene expression profile, epigenetic status, and self-renewal capacity. The e-iNSCs are functionally mature, as they could differentiate into all the neuronal cell types both in vitro and in vivo Our study provides a novel concept for generating functional iNSCs using a non-viral, non-integrating, plasmid-based system that could facilitate their biomedical applicability.
authors.affiliation: Department of Stem Cell Biology, School of Medicine, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea; Department of Animal Biotechnology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea.
authors.forename: Sung Min
authors.initials: SM
authors.lastname: Kim
authors.affiliation: Department of Nanobiomedical Science, Dankook University Graduate School, Cheonan 31116, Republic of Korea.
authors.forename: Jong-Wan
authors.initials: JW
authors.lastname: Kim
authors.affiliation: Department of Stem Cell Biology, School of Medicine, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea.
authors.forename: Tae Hwan
authors.initials: TH
authors.lastname: Kwak
authors.affiliation: Department of Physiology, School of Medicine, Konkuk University, Chungju, Chungbuk 27478, Republic of Korea.
authors.forename: Sang Woong
authors.initials: SW
authors.lastname: Park
authors.affiliation: Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstraße 20, 48149 Münster, Germany.
authors.forename: Kee-Pyo
authors.initials: KP
authors.lastname: Kim
authors.affiliation: Department of Physiology, School of Medicine, Konkuk University, Chungju, Chungbuk 27478, Republic of Korea.
authors.forename: Hyunji
authors.initials: H
authors.lastname: Park
authors.affiliation: Department of Stem Cell Biology, School of Medicine, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea.
authors.forename: Kyung Tae
authors.initials: KT
authors.lastname: Lim
authors.affiliation: Department of Stem Cell Biology, School of Medicine, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea.
authors.forename: Kyuree
authors.initials: K
authors.lastname: Kang
authors.affiliation: Department of Stem Cell Biology, School of Medicine, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea.
authors.forename: Jonghun
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstraße 20, 48149 Münster, Germany.
authors.forename: Ji Hun
authors.initials: JH
authors.lastname: Yang
authors.affiliation: Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 04056, Korea.
authors.forename: Heonjong
authors.initials: H
authors.lastname: Han
authors.affiliation: Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 04056, Korea.
authors.forename: Insuk
authors.initials: I
authors.lastname: Lee
authors.affiliation: Department of Nanobiomedical Science, Dankook University Graduate School, Cheonan 31116, Republic of Korea.
authors.forename: Jung Keun
authors.initials: JK
authors.lastname: Hyun
authors.affiliation: Department of Physiology, School of Medicine, Konkuk University, Chungju, Chungbuk 27478, Republic of Korea.
authors.forename: Young Min
authors.initials: YM
authors.lastname: Bae
authors.affiliation: Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstraße 20, 48149 Münster, Germany,; University of Münster, Medical Faculty, Domagkstraße 3, 48149 Münster, Germany.
authors.forename: Hans R
authors.initials: HR
authors.lastname: Schöler
authors.affiliation: Department of Animal Biotechnology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea.
authors.forename: Hoon Taek
authors.initials: HT
authors.lastname: Lee
authors.affiliation: Department of Stem Cell Biology, School of Medicine, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea; KU Open-Innovation Center, Institute of Biomedical Science & Technology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 05029, Republic of Korea. Electronic address: dwhan@konkuk.ac.kr.
authors.forename: Dong Wook
authors.initials: DW
authors.lastname: Han
mesh_terms: D000818:Animals; D005347:Fibroblasts / Q000166:cytology; D005822:Genetic Vectors; D000090062:Kruppel-Like Factor 4; D051379:Mice; D008809:Mice, Inbred C3H; D058953:Neural Stem Cells / Q000166:cytology*; D014162:Transfection
keywords: cell biology; cell therapy; neural stem cell (NSC); regenerative medicine; reprogramming