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Neurosci Res. 2016 Sep;110:18-28. doi: 10.1016/j.neures.2016.04.003. Epub 2016 Apr 13.

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.

Author information

1
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.
2
Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
3
Department of Orthopaedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
4
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.
5
Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-ku, Kanagawa, Japan.
6
Department of Orthopaedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan. Electronic address: masa@a8.keio.jp.
7
Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan. Electronic address: hidokano@a2.keio.jp.

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.

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

PMID:
27083781
DOI:
10.1016/j.neures.2016.04.003
[Indexed for MEDLINE]

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