title: In vitro detection of residual undifferentiated cells in retinal pigment epithelial cells derived from human induced pluripotent stem cells.
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.
authors.affiliation: Division of Cellular and Gene Therapy Products, National Institute of Health Sciences, 1-18-1 Kami-yoga, Setagaya, Tokyo, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Kuroda
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yasuda
authors.forename: Yoji
authors.initials: Y
authors.lastname: Sato
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D066298:In Vitro Techniques; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D061251:Primary Cell Culture; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction