Highly sensitive and non-disruptive detection of residual undifferentiated cells by measuring miRNAs in culture supernatant

Sci Rep. 2022 Jun 20;12(1):10351. doi: 10.1038/s41598-022-14273-z.

Abstract

The clinical usage of induced pluripotent stem cell (iPSC)-derived regenerative medicine products is limited by the possibility of residual undifferentiated cells forming tumours after transplantation. Most of the existing quality control tests involve crushing of cells. As a result, the cells to be transplanted cannot be directly tested, thereby increasing the cost of transplantation. Therefore, we tested a highly sensitive and non-disruptive quality-testing method that involves measuring microRNAs (miRNAs) in culture supernatants released by cells. By measuring miR-302b in the culture supernatant, residual iPSCs were detected with higher sensitivity than by measuring LIN28 (Lin-28 Homolog A) in the cells. To use this method, we also monitored the progression of differentiation. Our novel highly sensitive and non-disruptive method for detecting residual undifferentiated cells will contribute to reducing the manufacturing cost of iPSC-derived products and improving the safety of transplantation.

MeSH terms

  • Cell Differentiation
  • Induced Pluripotent Stem Cells*
  • MicroRNAs* / genetics

Substances

  • MicroRNAs