Development of selective cytotoxic viral vectors for concentration of undifferentiated cells in cardiomyocytes derived from human induced pluripotent stem cells

Ken Kono, Rumi Sawada, Takuya Kuroda, Satoshi Yasuda, Satoko Matsuyama, Akifumi Matsuyama, Hiroyuki Mizuguchi, Yoji Sato

Research output: Contribution to journalArticle

Abstract

Cell-processed therapeutic products (CTPs) derived from human pluripotent stem cells (hPSCs) have innovative applications in regenerative medicine. However, undifferentiated hPSCs possess tumorigenic potential; thus, sensitive methods for the detection of residual undifferentiated hPSCs are essential for the clinical use of hPSC-derived CTPs. The detection limit of the methods currently available is 1/10 5 (0.001%, undifferentiated hPSCs/differentiated cells) or more, which could be insufficient for the detection of residual hPSCs when CTPs contain more than 1 × 10 5 cells. In this study, we developed a novel approach to overcome this challenge, using adenovirus and adeno-associated virus (AdV and AAV)-based selective cytotoxic vectors. We constructed AdV and AAV vectors that possess a suicide gene, iCaspase 9 (iCasp9), regulated by the CMV promoter, which is dormant in hPSCs, for the selective expression of iCasp9 in differentiated cells. As expected, AdV/CMV-iCasp9 and AAV/CMV-iCasp9 exhibited cytotoxicity in cardiomyocytes but not in human induced pluripotent stem cells (hiPSCs). The vectors also induced apoptosis in hiPSC-derived cardiomyocytes, and the surviving cells exhibited higher levels of hPSC marker expression. These results indicate that the AdV- and AAV-based cytotoxic vectors concentrate cells expressing the undifferentiated cell markers in hiPSC-derived products and are promising biological tools for verifying the quality of CTPs.

Original languageEnglish
Article number3630
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 01-12-2019

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Induced Pluripotent Stem Cells
Pluripotent Stem Cells
Cardiac Myocytes
Dependovirus
Adenoviridae
Regenerative Medicine
Therapeutics
Suicide
Limit of Detection
Apoptosis

All Science Journal Classification (ASJC) codes

  • General

Cite this

Kono, Ken ; Sawada, Rumi ; Kuroda, Takuya ; Yasuda, Satoshi ; Matsuyama, Satoko ; Matsuyama, Akifumi ; Mizuguchi, Hiroyuki ; Sato, Yoji. / Development of selective cytotoxic viral vectors for concentration of undifferentiated cells in cardiomyocytes derived from human induced pluripotent stem cells. In: Scientific Reports. 2019 ; Vol. 9, No. 1.
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Development of selective cytotoxic viral vectors for concentration of undifferentiated cells in cardiomyocytes derived from human induced pluripotent stem cells. / Kono, Ken; Sawada, Rumi; Kuroda, Takuya; Yasuda, Satoshi; Matsuyama, Satoko; Matsuyama, Akifumi; Mizuguchi, Hiroyuki; Sato, Yoji.

In: Scientific Reports, Vol. 9, No. 1, 3630, 01.12.2019.

Research output: Contribution to journalArticle

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AU - Kono, Ken

AU - Sawada, Rumi

AU - Kuroda, Takuya

AU - Yasuda, Satoshi

AU - Matsuyama, Satoko

AU - Matsuyama, Akifumi

AU - Mizuguchi, Hiroyuki

AU - Sato, Yoji

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