RRC ID 64715
Author Mitsui K, Takahashi T, Ide K, Matsuda E, Kosai KI.
Title Optimization of adenoviral gene transfer in human pluripotent stem cells.
Journal Biochem Biophys Res Commun
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.
Volume 541
Pages 78-83
Published 2021-2-19
DOI 10.1016/j.bbrc.2021.01.009
PII S0006-291X(21)00042-5
PMID 33482579
MeSH Adenoviridae / genetics* Adenoviridae / physiology Cell Culture Techniques Cells, Cultured Gene Transfer Techniques / standards* Genes, Reporter / genetics Genetic Vectors / genetics* Green Fluorescent Proteins / genetics Humans Pluripotent Stem Cells / cytology Pluripotent Stem Cells / metabolism* Promoter Regions, Genetic / genetics
IF 2.985
Resource
DNA material pAxCAEGFP-Fwt (RDB03943) AxCAEGFP-F/RGD (RDB03010)
Human and Animal Cells 201B7(HPS0063)