RRC ID 64681
Author Watanabe T, Tateno H.
Title Elimination of cells deviated from human induced pluripotent stem cells with a photoactivatable IR700-labelled antibody.
Journal Biochem Biophys Res Commun
Abstract Human induced pluripotent stem cells (hiPSCs) are important starting materials for cell therapy products (CTPs) used for transplantation. During cell culture, hiPSCs often spontaneously undergo morphological changes and lose pluripotency. Such cells are called 'deviated cells', which are deviated from the undifferentiated state of hiPSCs, lack the expression of hiPSC markers and become positive for the early differentiation marker SSEA1 (stage-specific embryonic antigen 1, Lewis X glycan). Previously, we identified fibronectin (FN) as a predominant carrier protein of SSEA1 secreted from deviated cells, but not hiPSCs. A sandwich assay using antibodies (Abs) against FN and SSEA1 was developed for non-destructive quantitative evaluation of deviated cells present in hiPSC cultures. In this study, a novel technology was developed to specifically eliminate deviated cells using an anti-FN Ab along with a near-infrared (NIR) photoabsorber, IRDye700DX N-hydroxysuccinimide ester (IR700), which has been used for cancer photoimmunotherapy. The anti-FN Ab conjugated with the IR700 dye (IR700-αFN) bound to and induced the death of deviated cells upon NIR irradiation. In contrast, IR700-αFN failed to stain the hiPSCs, and IR700-αFN/NIR had little or no effect on survival. Finally, IR700-αFN/NIR irradiation induced selective removal of deviated cells from a mixed culture with hiPSCs, demonstrating that the proposed method is suitable for the removal of unwanted deviated cells present in hiPSC culture for the production of CTPs.
Volume 554
Pages 13-18
Published 2021-5-21
DOI 10.1016/j.bbrc.2021.03.078
PII S0006-291X(21)00469-1
PMID 33774274
MeSH Cell Culture Techniques Cell Proliferation Cell Separation / methods* Fibronectins / immunology Fibronectins / metabolism* Humans Immunoconjugates / pharmacology* Immunoconjugates / radiation effects Immunologic Factors / pharmacology Indoles / chemistry* Indoles / pharmacology Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / drug effects Infrared Rays Organosilicon Compounds / chemistry* Organosilicon Compounds / pharmacology
IF 2.985
Human and Animal Cells 201B7(HPS0063)