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Elimination of cells deviated from human induced pluripotent stem cells with a photoactivatable IR700-labelled antibody.
Watanabe, Tomoko; Tateno, Hiroaki.
Afiliación
  • Watanabe T; Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan.
  • Tateno H; Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. Electronic address: h-tateno@aist.go.jp.
Biochem Biophys Res Commun ; 554: 13-18, 2021 05 21.
Article en En | MEDLINE | ID: mdl-33774274
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Separación Celular / Fibronectinas / Compuestos de Organosilicio / Inmunoconjugados / Células Madre Pluripotentes Inducidas / Indoles Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Biochem Biophys Res Commun Año: 2021 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Separación Celular / Fibronectinas / Compuestos de Organosilicio / Inmunoconjugados / Células Madre Pluripotentes Inducidas / Indoles Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Biochem Biophys Res Commun Año: 2021 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos