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In Vitro Detection of Residual Undifferentiated Cells in Retinal Pigment Epithelial Cells Derived from Human Induced Pluripotent Stem Cells

  • Takuya Kuroda
  • Satoshi Yasuda
  • Yoji Sato
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1210)

Abstract

Human pluripotent stem cells (hPSCs) such as human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) are a leading candidate for regenerative medicine/cell therapies because of their capacity for pluripotency and unlimited self-renewal. However, there are significant obstacles preventing the clinical use of hPSCs. A significant safety issues is the presence of residual undifferentiated cells that have the potential to form tumors in vivo. Here, we describe the highly sensitive qRT-PCR methods for detection of residual undifferentiated cells in retinal pigment epithelial (RPE) cells derived from hiPSCs. qRT-PCR using probes and primers targeting LIN28A (LIN28) transcripts can detect residual undifferentiated cell levels as low as 0.002 % in hiPSC-derived RPE cells. We expect this method to contribute to process validation and quality control of hiPSC-derived cell therapy product.

Key words

Human induced pluripotent stem cells Retinal pigment epithelial cells Tumorigenicity LIN28 qRT-PCR 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Division of Cellular and Gene Therapy ProductsNational Institute of Health SciencesSetagayaJapan

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