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

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Stem Cells and Tissue Repair

Part of the book series: Methods in Molecular Biology ((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.

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Authors and Affiliations

  1. Division of Cellular and Gene Therapy Products, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo, Japan

    Takuya Kuroda, Satoshi Yasuda & Yoji Sato

Authors
  1. Takuya Kuroda
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  2. Satoshi Yasuda
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  3. Yoji Sato
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Corresponding author

Correspondence to Yoji Sato .

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Editors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, Oregon, USA

    Chrissa Kioussi

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Kuroda, T., Yasuda, S., Sato, Y. (2014). In Vitro Detection of Residual Undifferentiated Cells in Retinal Pigment Epithelial Cells Derived from Human Induced Pluripotent Stem Cells. In: Kioussi, C. (eds) Stem Cells and Tissue Repair. Methods in Molecular Biology, vol 1210. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1435-7_14

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  • DOI: https://doi.org/10.1007/978-1-4939-1435-7_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1434-0

  • Online ISBN: 978-1-4939-1435-7

  • eBook Packages: Springer Protocols

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