細胞・組織の高圧瞬間凍結法の開発

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2026-03-25 東京大学

東京大学と理化学研究所の研究チームは、大気圧の約2000倍の高圧下で数ミリ秒以内に細胞・組織を凍結する「高圧瞬間凍結法」を開発し、従来困難だった単層培養細胞や細胞凝集塊の凍結保存と高い生存率での再培養に成功した。電子顕微鏡用技術を応用し、毒性のある凍結保護剤を使わずに細胞活性を維持できる点が特徴である。本手法はオルガノイドやiPS細胞由来組織の保存に応用可能で、再生医療やバイオ研究の発展に貢献すると期待される。

細胞・組織の高圧瞬間凍結法の開発
細胞・組織の高圧瞬間凍結-実験の概要-

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凍結保存のための高圧凍結に関する概念実証研究 A proof-of-concept study on high pressure freezing for cryopreservation

Fang Song ,Mayuko Sato ,Yuya Toyama ,Taiyo Ishikawa ,Fumiya Tokito ,Takeshi Katsuda ,Kiminori Toyooka ,Yasuyuki Sakai ,Masaki Nishikawa
PNAS Nexus  Published:20 March 2026
DOI:https://doi.org/10.1093/pnasnexus/pgag065

Abstract

Cryopreservation by vitrification typically requires 30–50 v/v% of cytotoxic penetrable cryoprotective agents (CPAs) to prevent ice crystal formation during freezing and thawing, limiting its broader application. Since pressure suppresses ice crystallization, applying high pressure during vitrification may enable reducing CPA concentrations while maintaining cell viability. In this study, we used a high pressure freezing (HPF) device, commonly used for cryofixation, to successfully cryopreserve 2D cell monolayers and 3D cell spheroids with 20–30 v/v% penetrable CPA. Compared to commonly used plunge freezing, HPF cell monolayers exhibited higher post-thaw viability and better retention on the substrate, allowing for subsequent proliferation. HPF cell spheroids showed improved cell viability, metabolic activity and maintained cell-cell adhesion. Developing HPF devices specifically for cryopreservation, in combination with advanced warming techniques, holds promise for achieving vitrification with low or even no CPA.

生物化学工学
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