Formation of Large Scaffold-Free 3-D Aggregates in a Cell Culture Dish by Ultrasound Standing Wave Trapping

Misa Nakao, Chikahiro Imashiro, Taiki Kuribara, Yuta Kurashina, Kiichiro Totani, Kenjiro Takemura

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)


Cellular aggregates that mimic cell–cell interactions in vitro are essential for biological research. This study introduces a method to form large scaffold-free 3-D aggregates in a clinically ubiquitous cell culture dish using kilohertz-order ultrasound standing wave trapping (USWT). We fabricated an aggregate formation system in which a 60-mm dish was set above a Langevin transducer via water. The transducer was excited at 110.8 kHz, and then C2C12 myoblasts were injected into the dish and trapped at the node position of the standing wave. The diameter and thickness of the formed aggregate were 8 and 2.7 mm, respectively, which are larger than those of aggregates formed previously by USWT. Moreover, we confirmed that >94% of cells constituting the aggregates survived 9 h, and the protein expression of cells was not altered significantly. This method can be applied to form aggregates with high functionality, which contributes to the development of biological research methodology.

Original languageEnglish
Pages (from-to)1306-1315
Number of pages10
JournalUltrasound in Medicine and Biology
Issue number5
Publication statusPublished - 2019 May


  • Mouse myoblasts
  • Scaffold-free aggregates
  • Three-dimensional aggregates
  • Tissue engineering
  • Ultrasound standing wave trapping
  • Ultrasound transducer

ASJC Scopus subject areas

  • Radiological and Ultrasound Technology
  • Biophysics
  • Acoustics and Ultrasonics


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