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 journalArticle

5 Citations (Scopus)

Abstract

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 2μm, 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.

Original languageEnglish
JournalBiotechnology and Bioengineering
DOIs
Publication statusAccepted/In press - 2017

Fingerprint

Vibration
Cell culture
Vibrations (mechanical)
Enzymes
Modulation
Proteins
Membranes
Temperature
Thawing
Temperature control
Freezing
Trypsin
Membrane Proteins
Cells
Cell Culture Techniques
Myoblasts
Cell Count
Cell Line
Equipment and Supplies
Growth

Keywords

  • Cell culture
  • Cell detachment
  • Enzyme free
  • Myoblast
  • Resonance vibration
  • Temperature modulation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Enzyme-free cell detachment mediated by resonance vibration with temperature modulation. / Kurashina, Yuta; Hirano, Makoto; Imashiro, Chikahiro; Totani, Kiichiro; Komotori, Jun; Takemura, Kenjiro.

In: Biotechnology and Bioengineering, 2017.

Research output: Contribution to journalArticle

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AU - Komotori, Jun

AU - Takemura, Kenjiro

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