An effective detachment system for human induced pluripotent stem cells cultured on multilayered cultivation substrates using resonance vibrations

Yusuke Terao, Yuta Kurashina, Shugo Tohyama, Yuki Fukuma, Keiichi Fukuda, Jun Fujita, Kenjiro Takemura

Research output: Contribution to journalArticle

Abstract

Clinical application of human induced pluripotent stem cells (hiPSCs) has been hampered by the lack of a practical, scalable culture system. Stacked culture plates (SCPs) have recently attracted attention. However, final cell yields depend on the efficiency of cell detachment, and inefficient cell recovery from SCPs presents a major challenge to their use. We have developed an effective detachment method using resonance vibrations (RVs) of substrates with sweeping driving frequency. By exciting RVs that have 1–3 antinodes with ultra-low-density enzyme spread on each substrate of SCPs, 87.8% of hiPSCs were successfully detached from a 5-layer SCP compared to 30.8% detached by the conventional enzymatic method. hiPSC viability was similar after either method. Moreover, hiPSCs detached by the RV method maintained their undifferentiated state. Additionally, hiPSCs after long-term culture (10 passages) kept excellent detachment efficiency, had the normal karyotypes, and maintained the undifferentiated state and pluripotency. These results indicated that the RV method has definite advantages over the conventional enzymatic method in the scalable culture of hiPSCs using SCPs.

Original languageEnglish
Article number15655
JournalScientific reports
Volume9
Issue number1
DOIs
Publication statusPublished - 2019 Dec 1

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Induced Pluripotent Stem Cells
Vibration
Karyotype
Enzymes

ASJC Scopus subject areas

  • General

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An effective detachment system for human induced pluripotent stem cells cultured on multilayered cultivation substrates using resonance vibrations. / Terao, Yusuke; Kurashina, Yuta; Tohyama, Shugo; Fukuma, Yuki; Fukuda, Keiichi; Fujita, Jun; Takemura, Kenjiro.

In: Scientific reports, Vol. 9, No. 1, 15655, 01.12.2019.

Research output: Contribution to journalArticle

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AU - Fukuda, Keiichi

AU - Fujita, Jun

AU - Takemura, Kenjiro

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