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Please use this identifier to cite or link to this item: http://hdl.handle.net/10119/13700

Title: Cryopreservation of a two-dimensional monolayer using a slow vitrification method with polyampholyte to inhibit ice crystal formation
Authors: Matsumura, Kazuaki
Kawamoto, Keiko
Takeuchi, Masahiro
Yoshimura, Shigehiro
Tanaka, Daisuke
Hyon, Suong-Hyu
Keywords: cryopreservation
ice crystallization
polyampholyte
tissue-engineered construct
vitrification
Issue Date: 2016-04-18
Publisher: American Chemical Society
Magazine name: ACS Biomaterials Science & Engineering
Volume: 2
Number: 6
Start page: 1023
End page: 1029
DOI: 10.1021/acsbiomaterials.6b00150
Abstract: Vitrification methods have been developed to improve the preservation of oocytes and embryos. However, successful vitrification and preservation typically requires very high cooling speeds. Here, we report a novel slow vitrification method for cryopreservation of two-dimensional (2D) cell constructs using a vitrification solution (VS) in PBS containing 6.5 M ethylene glycol, 0.5 M sucrose, and 10% w/w carboxylated poly-L-lysine (COOH-PLL), a novel polymeric cryoprotectant and stabilizing agent that is likely to inhibit ice crystallization. Stabilization of the glassy state and inhibition of devitrification was confirmed by thermal analysis using differential scanning calorimetry. The viability of cultured human mesenchymal stem cell (MSC) monolayers after freezing by our novel slow vitrification method at a rate of 4.9°C/min in VS with 10% COOH-PLL was significantly higher than that of cells frozen using our slow vitrification method in VS without COOH-PLL. Moreover, cells maintained the capacity for differentiation. We further confirmed that COOH-PLL improved the vitrification properties of the current vitrification system through inhibition of recrystallization properties. This novel, simple method for slow vitrification can be widely applicable for the preservation of tissue-engineered constructs and may facilitate the industrialization of regenerative medicine.
Rights: Kazuaki Matsumura, Keiko Kawamoto, Masahiro Takeuchi, Shigehiro Yoshimura, Daisuke Tanaka, and Suong-Hyu Hyon, ACS Biomaterials Science & Engineering, 2016, 2(6), pp.1023-1029. This document is the unedited author's version of a Submitted Work that was subsequently accepted for publication in ACS Biomaterials Science & Engineering, copyright (c) American Chemical Society after peer review. To access the final edited and published work, see http://dx.doi.org/10.1021/acsbiomaterials.6b00150
URI: http://hdl.handle.net/10119/13700
Material Type: author
Appears in Collections:c10-1. 雑誌掲載論文 (Journal Articles)

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