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Please use this identifier to cite or link to this item: http://hdl.handle.net/10119/11595

Title: Development of a novel vitrification method for chondrocyte sheets
Authors: Maehara, Miki
Sato, Masato
Watanabe, Masahito
Matsunari, Hitomi
Kokubo, Mami
Kanai, Takahiro
Sato, Michio
Matsumura, Kazuaki
Hyon, Suong-Hyu
Yokoyama, Munetaka
Mochida, Joji
Nagashima, Hiroshi
Keywords: cell sheet
chondrocyte shee
tissue engineering
Issue Date: 2013-07-25
Publisher: BioMed Central Ltd.
Magazine name: BMC Biotechnology
Volume: 13
Start page: article no. 58
DOI: 10.1186/1472-6750-13-58
Abstract: Background: There is considerable interest in using cell sheets for the treatment of various lesions as part of regenerative medicine therapy. Cell sheets can be prepared in temperature-responsive culture dishes and applied to injured tissue. For example, cartilage-derived cell sheets are currently under preclinical testing for use in treatment of knee cartilage injuries. The additional use of cryopreservation technology could increase the range and practicality of cell sheet therapies. To date, however, cryopreservation of cell sheets has proved impractical. Results: Here we have developed a novel and effective method for cryopreserving fragile chondrocyte sheets. We modified the vitrification method previously developed for cryopreservation of mammalian embryos to vitrify a cell sheet through use of a minimum volume of vitrification solution containing 20% dimethyl sulfoxide, 20% ethylene glycol, 0.5 M sucrose, and 10% carboxylated poly-L-lysine. The principal feature of our method is the coating of the cell sheet with a viscous vitrification solution containing permeable and non-permeable cryoprotectants prior to vitrification in liquid nitrogen vapor. This method prevented fracturing of the fragile cell sheet even after vitrification and rewarming. Both the macro- and microstructures of the vitrified cell sheets were maintained without damageor loss of major components. Cell survival in the vitrified sheets was comparable to that in non-vitrified samples. Conclusions: We have shown here that it is feasible to vitrify chondrocyte cell sheets and that these sheets retain their normal characteristics upon thawing. The availability of a practical cryopreservation method should make a significant contribution to the effectiveness and range of applications of cell sheet therapy.
Rights: © 2013 Maehara et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Miki Maehara, Masato Sato, Masahito Watanabe, Hitomi Matsunari, Mami Kokubo, Takahiro Kanai, Michio Sato, Kazuaki Matsumura, Suong-Hyu Hyon, Munetaka Yokoyama, Joji Mochida and Hiroshi Nagashima, BMC Biotechnology, 13, 2013, article no. 58. http://dx.doi.org/10.1186/1472-6750-13-58
URI: http://hdl.handle.net/10119/11595
Material Type: publisher
Appears in Collections:c10-1. 雑誌掲載論文 (Journal Articles)

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