A Selective Cytotoxic Adenovirus Vector for Concentration of Pluripotent Stem Cells in Human Pluripotent Stem Cell-derived Neural Progenitor Cells

Overview

Journal Sci Rep

Specialty Science

Date 2021 Jun 2

PMID 34075124

Citations 1

Authors

Takamasa Hirai

Ken Kono

Rumi Sawada

Takuya Kuroda

Satoshi Yasuda

Satoko Matsuyama

Akifumi Matsuyama

Naoya Koizumi

Naoki Utoguchi

Hiroyuki Mizuguchi

Yoji Sato

Affiliations

Soon will be listed here.

Abstract

Highly sensitive detection of residual undifferentiated pluripotent stem cells is essential for the quality and safety of cell-processed therapeutic products derived from human induced pluripotent stem cells (hiPSCs). We previously reported the generation of an adenovirus (Ad) vector and adeno-associated virus vectors that possess a suicide gene, inducible Caspase 9 (iCasp9), which makes it possible to sensitively detect undifferentiated hiPSCs in cultures of hiPSC-derived cardiomyocytes. In this study, we investigated whether these vectors also allow for detection of undifferentiated hiPSCs in preparations of hiPSC-derived neural progenitor cells (hiPSC-NPCs), which have been expected to treat neurological disorders. To detect undifferentiated hiPSCs, the expression of pluripotent stem cell markers was determined by immunostaining and flow cytometry. Using immortalized NPCs as a model, the Ad vector was identified to be the most efficient among the vectors tested in detecting undifferentiated hiPSCs. Moreover, we found that the Ad vector killed most hiPSC-NPCs in an iCasp9-dependent manner, enabling flow cytometry to detect undifferentiated hiPSCs intermingled at a lower concentration (0.002%) than reported previously (0.1%). These data indicate that the Ad vector selectively eliminates hiPSC-NPCs, thus allowing for sensitive detection of hiPSCs. This cytotoxic viral vector could contribute to ensuring the quality and safety of hiPSCs-NPCs for therapeutic use.

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