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

doi:10.1016/j.ultrasmedbio.2019.01.013

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

Ultrasound Med Biol. 2019 May;45(5):1306-1315. doi: 10.1016/j.ultrasmedbio.2019.01.013. Epub 2019 Feb 22.

Authors

Misa Nakao¹, Chikahiro Imashiro¹, Taiki Kuribara², Yuta Kurashina³, Kiichiro Totani², Kenjiro Takemura⁴

Affiliations

¹ School of Science for Open and Environmental Systems, Graduate School of Science and Technology, Keio University, Yokohama, Kanagawa, Japan.
² Department of Materials and Life Science, Faculty of Science and Technology, Seikei University, Musashino, Tokyo, Japan.
³ Department of Materials Science and Engineering, School of Materials and Chemical Technology, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan 226-8503; Department of Mechanical Engineering, Faculty of Science and Technology, Keio University, Yokohama, Kanagawa, Japan.
⁴ Department of Mechanical Engineering, Faculty of Science and Technology, Keio University, Yokohama, Kanagawa, Japan. Electronic address: takemura@mech.keio.ac.jp.

PMID: 30799124
DOI: 10.1016/j.ultrasmedbio.2019.01.013

Abstract

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.

Keywords: Mouse myoblasts; Scaffold-free aggregates; Three-dimensional aggregates; Tissue engineering; Ultrasound standing wave trapping; Ultrasound transducer.

MeSH terms

Animals
Cell Aggregation / physiology
Cell Culture Techniques / methods*
Mice
Models, Animal
Myoblasts / physiology*
Tissue Engineering / methods*
Ultrasonic Waves*