title: Robust production of human neural cells by establishing neuroepithelial-like stem cells from peripheral blood mononuclear cell-derived feeder-free iPSCs under xeno-free conditions.
abstract: Neural stem/progenitor cells (NS/PCs) derived from human induced pluripotent stem cells (hiPSCs) are expected to be a valuable cell source for cell therapies that target central nervous system disorders. For clinical applications, NS/PCs should be induced and maintained under clinical grade conditions, which are challenging to achieve. In the present study, we established a procedure to obtain xeno-free long-term self-renewing neuroepithelial-like stem cells (xf-lt-NES cells) from feeder-free hiPSCs using a newly developed xeno-free medium, StemFit(®)AS200. xf-lt-NES cells were cultured for long periods in StemFit(®)AS200 while retaining normal karyotypes, NS/PC marker expression and differentiation capacity for neuronal and glial differentiation in vitro and in vivo. Furthermore, the cells were cryopreserved using a defined serum-free freezing reagent, which demonstrated the feasibility of this xeno-free culture system for large-scale lt-NES cell production and cell banking. Taken together, our system represents a promising approach for the manufacture of clinically relevant products for cell therapy using NS/PCs.
authors.affiliation: Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; Regenerative & Cellular Medicine Office, Sumitomo Dainippon Pharma Co., Ltd., Chuo-ku, Kobe, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Isoda
authors.affiliation: Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kohyama
authors.affiliation: Department of Orthopaedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Iwanami
authors.affiliation: Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; Department of Orthopaedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Sanosaka
authors.affiliation: Department of Orthopaedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Sugai
authors.affiliation: Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; Regenerative & Cellular Medicine Office, Sumitomo Dainippon Pharma Co., Ltd., Chuo-ku, Kobe, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Yamaguchi
authors.affiliation: Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-ku, Kanagawa, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Matsumoto
authors.affiliation: Department of Orthopaedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan. Electronic address: masa@a8.keio.jp.
authors.forename: Masaya
authors.initials: M
authors.lastname: Nakamura
authors.affiliation: Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan. Electronic address: hidokano@a2.keio.jp.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Okano
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D003470:Culture Media; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology* / Q000637:transplantation; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology
chemical_list: D003470:Culture Media
keywords: Long-term self-renewing neuroepithelial-like stem cells (lt-NES cells); Neural induction; Peripheral blood mononuclear cell-derived feeder-free human iPSCs; Regenerative medicine; Xeno-free