Seamless and non-destructive monitoring of extracellular microRNAs during cardiac differentiation from human pluripotent stem cells

Otoya Sekine, Sayaka Kanaami, Kanako Masumoto, Yuki Aihara, Yuika Morita-Umei, Hidenori Tani, Yusuke Soma, Tomohiko C. Umei, Kotaro Haga, Taijun Moriwaki, Yujiro Kawai, Masatoshi Ohno, Yoshikazu Kishino, Hideaki Kanazawa, Keiichi Fukuda, Masaki Ieda, Shugo Tohyama

研究成果: ジャーナルへの寄稿学術論文査読

1 被引用数 (Scopus)

抄録

Monitoring cardiac differentiation and maturation from human pluripotent stem cells (hPSCs) and detecting residual undifferentiated hPSCs are indispensable for the development of cardiac regenerative therapy. MicroRNA (miRNA) is secreted from cells into the extracellular space, and its role as a biomarker is attracting attention. Here, we performed an miRNA array analysis of supernatants during the process of cardiac differentiation and maturation from hPSCs. We demonstrated that the quantification of extracellular miR-489-3p and miR-1/133a-3p levels enabled the monitoring of mesoderm and cardiac differentiation, respectively, even in clinical-grade mass culture systems. Moreover, extracellular let-7c-5p levels showed the greatest increase with cardiac maturation during long-term culture. We also verified that residual undifferentiated hPSCs in hPSC-derived cardiomyocytes (hPSC-CMs) were detectable by measuring miR-302b-3p expression, with a detection sensitivity of 0.01%. Collectively, we demonstrate that our method of seamlessly monitoring specific miRNAs secreted into the supernatant is non-destructive and effective for the quality evaluation of hPSC-CMs.

本文言語英語
ページ(範囲)1925-1939
ページ数15
ジャーナルStem Cell Reports
18
10
DOI
出版ステータス出版済み - 10-10-2023
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生化学
  • 遺伝学
  • 発生生物学
  • 細胞生物学

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