イマシロ　チカヒロ   Imashiro Chikahiro   今城　哉裕    所属   医学研究科 医学研究科 (医学部医学科をご参照ください)    職種   非常勤講師
論文種別	原著
言語種別	英語
査読の有無	査読あり
表題	Enzyme-free cell detachment mediated by resonance vibration with temperature modulation.
掲載誌名	正式名：Biotechnology and bioengineering 略　 称：Biotechnol Bioeng ISSNコード：10970290/00063592
掲載区分	国外
巻・号・頁	114(10),pp.2279-2288
著者・共著者	Kurashina Yuta†, Hirano Makoto, Imashiro Chikahiro, Totani Kiichiro, Komotori Jun, Takemura Kenjiro*
発行年月	2017/10
概要	Cell detachment is an essential process in adherent cell culture. However, trypsinization, which is the most popular detachment technique used in culture, damages cellular membranes. Reducing cellular membrane damage during detachment should improve the quality of cell culture. In this article, we propose an enzyme-free cell detachment method based on resonance vibration with temperature modulation. We developed a culture device that can excite a resonance vibration and control temperature. We then evaluated the cell detachment ratio and the growth response, observed the morphology, and analyzed the cellular protein of the collected cells-mouse myoblast cell line (C2C12). With the temperature of 10°C and the maximum vibration amplitude of 2 μm, 77.9% of cells in number were successfully detached compared with traditional trypsinization. The 72-h proliferation ratio of the reseeded cells was similar to that with trypsinization, whereas the proliferation ratio of proposed method was 12.6% greater than that of trypsinization after freezing and thawing. Moreover, the cells can be collected relatively intact and both intracellular and cell surface proteins in the proposed method were less damaged than in trypsinization. These results show that this method has definite advantages over trypsinization, which indicates that it could be applied to subcultures of cells that are more susceptible to trypsin damage for mass culture of sustainable clinical use. Biotechnol. Bioeng. 2017;114: 2279-2288. © 2017 Wiley Periodicals, Inc.
DOI	10.1002/bit.26361
PMID	28627736