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Optimization of adenoviral gene transfer in human pluripotent stem cells.
Mitsui, Kaoru; Takahashi, Tomoyuki; Ide, Kanako; Matsuda, Eriko; Kosai, Ken-Ichiro.
Afiliação
  • Mitsui K; Department of Gene Therapy and Regenerative Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544, Japan; Center for Innovative Therapy Research and Application, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544, Jap
  • Takahashi T; Department of Pediatrics and Child Health, Kurume University School of Medicine, Fukuoka, 830-0011, Japan; Cognitive and Molecular Research Institute of Brain Diseases, Kurume University, Fukuoka, 830-0011, Japan.
  • Ide K; Department of Gene Therapy and Regenerative Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544, Japan.
  • Matsuda E; Department of Gene Therapy and Regenerative Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544, Japan.
  • Kosai KI; Department of Gene Therapy and Regenerative Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544, Japan; Center for Innovative Therapy Research and Application, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544, Jap
Biochem Biophys Res Commun ; 541: 78-83, 2021 02 19.
Article em En | MEDLINE | ID: mdl-33482579
ABSTRACT
Human pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells, have the potential to differentiate into a wide variety of cells in vitro and have applications in basic developmental biology research and regenerative medicine. To understand the process of differentiation from pluripotent stem cells to functional cells, it is necessary to efficiently and safely transfer and express exogenous genes. We attempted to optimize the efficient transfer of genes into pluripotent stem cells using adenoviral vectors. Comparative study of the activities of three representative ubiquitously active promoters revealed that only the CA promoter allowed robust transgene expression in human pluripotent stem cells. In addition, we established a protocol that allowed us to efficiently introduce target genes and ensure their expression even in small numbers of cells. Adenoviral vector infection of pluripotent stem cells in single-cell suspension culture yielded high gene transfer efficiency with low cytotoxicity, without losing the undifferentiated state of the pluripotent stem cells. This optimized system will facilitate developmental biology research and regenerative medicine using pluripotent stem cells.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Adenoviridae / Técnicas de Transferência de Genes / Células-Tronco Pluripotentes / Vetores Genéticos Limite: Humans Idioma: En Revista: Biochem Biophys Res Commun Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Adenoviridae / Técnicas de Transferência de Genes / Células-Tronco Pluripotentes / Vetores Genéticos Limite: Humans Idioma: En Revista: Biochem Biophys Res Commun Ano de publicação: 2021 Tipo de documento: Article
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