Enzyme-free cell detachment mediated by resonance vibration with temperature modulation

Yuta Kurashina, Makoto Hirano, Chikahiro Imashiro, Kiichiro Totani, Jun Komotori, Kenjiro Takemura

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)2279-2288
Number of pages10
JournalBiotechnology and bioengineering
Issue number10
Publication statusPublished - 2017 Oct


  • cell culture
  • cell detachment
  • enzyme free
  • myoblast
  • resonance vibration
  • temperature modulation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


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