Yuta Tsujisaka,^1,2,5 Takeshi Hatani,^1,2,5 Chikako Okubo,¹ Ryo Ito,¹ Azuma Kimura,¹ Megumi Narita,¹

Kazuhisa Chonabayashi,^1,3 Shunsuke Funakoshi,^1,4 Antonio Lucena-Cacace,¹ Taro Toyoda,1 Kenji Osafune,¹

Takeshi Kimura,² Hirohide Saito,^{1, *} and Yoshinori Yoshida^{1,4, *}

¹Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan

²Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan

³Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan

⁴Takeda-CiRA Joint Program (T-CiRA), Fujisawa 251-0012, Japan

⁵These authors contributed equally

*Correspondence: 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。 (H.S.), 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。 (Y.Y.)

https://doi.org/10.1016/j.stemcr.2022.05.003

SUMMARY
For regenerative cell therapies using pluripotent stem cell (PSC)-derived cells, large quantities of purified cells are required. Magnetic-activated cell sorting (MACS) is a powerful approach to collect target antigen-positive cells; however, it remains a challenge to purify various cell types efficiently at large scale without using antibodies specific to the desired cell type. Here we develop a technology that combines microRNA (miRNA)-responsive mRNA switch (miR-switch) with MACS (miR-switch-MACS) to purify large amounts of PSC-derived cells rapidly and effectively. We designed miR-switches that detect specific miRNAs expressed in target cells and controlled the translation of a CD4-coding transgene as a selection marker for MACS. For the large-scale purification of induced PSC-derived cardiomyocytes (iPSC-CMs), we transferred miR-208a-CD4 switch-MACS and obtained purified iPSC-CMs efficiently. Moreover, miR-375-CD4 switch-MACS highly purified pancreatic insulin-producing cells and their progenitors expressing Chromogranin A. Overall, the miR-switch-MACS method can efficiently purify target PSC-derived cells for cell replacement therapy.