Cryopreservation of undifferentiated and differentiated human neuronal cells

Kenji Yamatoya; Yuya Nagai; Naozumi Teramoto; Woojin Kang; Kenji Miyado; Kazuya Nakata; Tohru Yagi; Yoshitaka Miyamoto

Regenerative Therapy (Mar 2022)

Cryopreservation of undifferentiated and differentiated human neuronal cells

Kenji Yamatoya,
Yuya Nagai,
Naozumi Teramoto,
Woojin Kang,
Kenji Miyado,
Kazuya Nakata,
Tohru Yagi,
Yoshitaka Miyamoto

Affiliations

Kenji Yamatoya: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan; Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Tomioka, Urayasu, Chiba, 279-0021, Japan
Yuya Nagai: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan
Naozumi Teramoto: Department of Applied Chemistry, Faculty of Engineering, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino, Chiba, 275-0016, Japan
Woojin Kang: Department of Reproductive Biology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo, 157-8535, Japan
Kenji Miyado: Department of Reproductive Biology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo, 157-8535, Japan
Kazuya Nakata: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan; Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Nakacho, Koganei, Tokyo, 184-0012, Japan
Tohru Yagi: Department of Mechanical Engineering, Tokyo Institute of Technology, 12-2-1 Ookayama, Meguro-ku, Tokyo, 152-8552, Japan
Yoshitaka Miyamoto: Department of Applied Chemistry, Faculty of Engineering, Chiba Institute of Technology, 2-17-1 Tsudanuma, Narashino, Chiba, 275-0016, Japan; Department of Reproductive Biology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo, 157-8535, Japan; Department of Mechanical Engineering, Tokyo Institute of Technology, 12-2-1 Ookayama, Meguro-ku, Tokyo, 152-8552, Japan; Corresponding author. Department of Reproductive Biology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo, 157-8535, Japan. Fax: +81-3-5494-7048.

Journal volume & issue: Vol. 19
pp. 58 – 68

Abstract

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The effective use of human-derived cells that are difficult to freeze, such as parenchymal cells and differentiated cells from stem cells, is crucial. A stable supply of damage-sensitive cells, such as differentiated neuronal cells, neurons, and glial cells can contribute considerably to cell therapy. We developed a serum-free freezing solution that is effective for the cryopreservation of differentiated neuronal cells. The quality of the differentiated and undifferentiated SK-N-SH cells was determined based on cell viability, live-cell recovery rate, and morphology of cultured cells, to assess the efficacy of the freezing solutions. The viability and recovery rate of the differentiated SK-N-SH neuronal cells were reduced by approximately 1.5-folds compared to that of the undifferentiated SK-N-SH cells. The viability and recovery rate of the differentiated SK-N-SH cells were remarkably different between the freezing solutions containing 10% DMSO and that containing 10% glycerol. Cryoprotectants such as fetal bovine serum (FBS), antifreeze proteins (sericin), and sugars (maltose), are essential for protecting against freeze damage in differentiated neuronal cells and parenchymal cells. Serum-free alternatives (sericin and maltose) could increase safety during cell transplantation and regenerative medicine. Considering these, we propose an effective freezing solution for the cryopreservation of neuronal cells.

Published in Regenerative Therapy

ISSN: 2352-3204 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Medicine: Medicine (General); Science: Biology (General): Cytology
Website: http://www.journals.elsevier.com/regenerative-therapy/

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