TY - JOUR
T1 - Safe and efficient method for cryopreservation of human induced pluripotent stem cell-derived neural stem and progenitor cells by a programmed freezer with a magnetic field
AU - Nishiyama, Yuichiro
AU - Iwanami, Akio
AU - Kohyama, Jun
AU - Itakura, Go
AU - Kawabata, Soya
AU - Sugai, Keiko
AU - Nishimura, Soraya
AU - Kashiwagi, Rei
AU - Yasutake, Kaori
AU - Isoda, Miho
AU - Matsumoto, Morio
AU - Nakamura, Masaya
AU - Okano, Hideyuki
N1 - Funding Information:
We thank Prof. Yoshiaki Toyama (Keio University) for invaluable comments, Prof. Shinya Yamanaka (Kyoto University) for the undifferentiated iPSCs, Prof. Douglass Sipp (Keio University) for invaluable comments for the manuscript, Prof. Robert Darnell (Rockefeller University) for the anti-Hu antibody and Mr. Owada (ABI corporation Ltd., Abiko, Japan) for the CAS. We also thank Drs. F. Renault-Mihara, M. Shinozaki, S. Shibata, R. Yamaguchi, A. Yasuda, S. Nori, Y. Kobayashi, M. Takano, T. Konomi, H. Iwai, S. Tashiro, M. Ozaki, T. Iida, K. Matsubayashi, T. Okubo, K. Kojima, S. Ito and Ms. Harada, who are the members of the spinal cord research team in the Departments of Orthopedic Surgery, Rehabilitation Medicine, and Physiology in Keio University School of Medicine for their assistance. This work was supported by Research Center Network for Realization of Regenerative Medicine by the Japan Science and Technology Agency (JST) and the Japan Agency for Medical Research and Development (AMED) (to H.O. and M.N.). This work was also supported by the Research Project for Practical Application of Regenerative Medicine from AMED (to A.I., J.K. and M.N.). This work was carried out in collaboration with Keio University School of Medicine, ABI Corporation Ltd., and Nihon Unisys Ltd. H.O. is a paid scientific advisory board member for SanBio Co., Ltd.
Publisher Copyright:
© 2015 The Authors.
PY - 2016/6/1
Y1 - 2016/6/1
N2 - Stem cells represent a potential cellular resource in the development of regenerative medicine approaches to the treatment of pathologies in which specific cells are degenerated or damaged by genetic abnormality, disease, or injury. Securing sufficient supplies of cells suited to the demands of cell transplantation, however, remains challenging, and the establishment of safe and efficient cell banking procedures is an important goal. Cryopreservation allows the storage of stem cells for prolonged time periods while maintaining them in adequate condition for use in clinical settings. Conventional cryopreservation systems include slow-freezing and vitrification both have advantages and disadvantages in terms of cell viability and/or scalability. In the present study, we developed an advanced slow-freezing technique using a programmed freezer with a magnetic field called Cells Alive System (CAS) and examined its effectiveness on human induced pluripotent stem cell-derived neural stem/progenitor cells (hiPSC-NS/PCs). This system significantly increased cell viability after thawing and had less impact on cellular proliferation and differentiation. We further found that frozen-thawed hiPSC-NS/PCs were comparable with non-frozen ones at the transcriptome level. Given these findings, we suggest that the CAS is useful for hiPSC-NS/PCs banking for clinical uses involving neural disorders and may open new avenues for future regenerative medicine.
AB - Stem cells represent a potential cellular resource in the development of regenerative medicine approaches to the treatment of pathologies in which specific cells are degenerated or damaged by genetic abnormality, disease, or injury. Securing sufficient supplies of cells suited to the demands of cell transplantation, however, remains challenging, and the establishment of safe and efficient cell banking procedures is an important goal. Cryopreservation allows the storage of stem cells for prolonged time periods while maintaining them in adequate condition for use in clinical settings. Conventional cryopreservation systems include slow-freezing and vitrification both have advantages and disadvantages in terms of cell viability and/or scalability. In the present study, we developed an advanced slow-freezing technique using a programmed freezer with a magnetic field called Cells Alive System (CAS) and examined its effectiveness on human induced pluripotent stem cell-derived neural stem/progenitor cells (hiPSC-NS/PCs). This system significantly increased cell viability after thawing and had less impact on cellular proliferation and differentiation. We further found that frozen-thawed hiPSC-NS/PCs were comparable with non-frozen ones at the transcriptome level. Given these findings, we suggest that the CAS is useful for hiPSC-NS/PCs banking for clinical uses involving neural disorders and may open new avenues for future regenerative medicine.
KW - Allogeneic transplantation
KW - Cells Alive System (CAS)
KW - Central nervous system (CNS) disorder
KW - Cryopreservation
KW - Human iPSC-derived neural stem/progenitor cells (hiPSC-NS/PCs)
KW - Magnetic field
KW - Neurosphere
KW - Spinal cord injury (SCI)
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U2 - 10.1016/j.neures.2015.11.011
DO - 10.1016/j.neures.2015.11.011
M3 - Article
C2 - 26804710
AN - SCOPUS:84960145049
SN - 0168-0102
VL - 107
SP - 20
EP - 29
JO - Neuroscience Research
JF - Neuroscience Research
ER -