MicroRNA-302 switch to identify and eliminate undifferentiated human pluripotent stem cells

Callum J.C. Parr, Shota Katayama, Kenji Miki, Yi Kuang, Yoshinori Yoshida, Asuka Morizane, Jun Takahashi, Shinya Yamanaka, Hirohide Saito

Research output: Contribution to journalArticlepeer-review

40 Citations (Scopus)

Abstract

The efficiency of pluripotent stem cell differentiation is highly variable, often resulting in heterogeneous populations that contain undifferentiated cells. Here we developed a sensitive, target-specific, and general method for removing undesired cells before transplantation. MicroRNA-302a-5p (miR-302a) is highly and specifically expressed in human pluripotent stem cells and gradually decreases to basal levels during differentiation. We synthesized a new RNA tool, miR-switch, as a live-cell reporter mRNA for miR-302a activity that can specifically detect human induced pluripotent stem cells (hiPSCs) down to a spiked level of 0.05% of hiPSCs in a heterogeneous population and can prevent teratoma formation in an in vivo tumorigenicity assay. Automated and selective hiPSC-elimination was achieved by controlling puromycin resistance using the miR-302a switch. Our system uniquely provides sensitive detection of pluripotent stem cells and partially differentiated cells. In addition to its ability to eliminate undifferentiated cells, miR-302a switch also holds great potential in investigating the dynamics of differentiation and/or reprograming of live-cells based on intracellular information.

Original languageEnglish
Article number32532
JournalScientific reports
Volume6
DOIs
Publication statusPublished - 2016 Sep 9

ASJC Scopus subject areas

  • General

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