Optimization of Zwitterionic Polymers for Cell Cryopreservation

Author:

Kato Yui1,Uto Takuya2,Ishizaki Takeru1,Tanaka Daisuke3,Ishibashi Kojiro4,Matsuda Yuya1,Onoda Issei2,Kobayashi Akiko5,Hazawa Masaharu5,Wong Richard W.5,Takahashi Kenji1,Hirata Eishu46,Kuroda Kosuke1ORCID

Affiliation:

1. Faculty of Biological Science and Technology Institute of Science and Engineering Kanazawa University Kakuma‐machi Kanazawa 920‐1192 Japan

2. University of Miyazaki Faculty of Engineering Nishi 1‐1 Gakuen Kibanadai Miyazaki 889‐2192 Japan

3. Research Center of Genetic Resources National Agriculture and Food Research Organization Kannondai Tsukuba 305‐8602 Japan

4. Cancer Research Institute Kanazawa University Kakuma‐machi Kanazawa 920‐1192 Japan

5. Cell‐Bionomics Research Unit Institute for Frontier Science Initiative & WPI‐Nano Life Science Institute Kanazawa University, Kanazawa Kanazawa Ishikawa 920‐1192 Japan

6. Nano Life Science Institute of Kanazawa University Kakuma‐machi Kanazawa 920‐1192 Japan

Abstract

AbstractCryopreservation techniques are valuable for the preservation of genetic properties in cells, and the development of this technology contributes to various fields. In a previous study, an isotonic freezing medium composed of poly(zwitterion) (polyZI) has been reported, which alleviates osmotic shock, unlike typical hypertonic freezing media. In this study, the primitive freezing medium composed of emerging polyZI is optimized. Imidazolium/carboxylate‐type polyZI (VimC3C) is the optimal chemical structure. The molecular weight and degree of ion substitution (DSion) are not significant factors. There is an impediment with the primitive polyZI freezing media. While the polyZI forms a matrix around the cell membrane to protect cells, the matrix is difficult to remove after thawing, resulting in low cell proliferation. Unexpectedly, increasing the poly(VimC3C) concentration from 10% to 20% (w/v) improves cell proliferation. The optimized freezing medium, 20% (w/v) poly(VimC3C)_DSion(100%)/1% (w/v) NaCl aqueous solution, exhibited a better cryoprotective effect.

Funder

Yamaguchi University

National Institute for Basic Biology

Kanazawa University

Japan Science and Technology Agency

ACT-X

Japan Society for the Promotion of Science

Publisher

Wiley

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