Optimization of Adenoviral Gene Transfer in Human Pluripotent Stem Cells
Optimization of Adenoviral Gene Transfer in Human Pluripotent Stem Cells
复制标题
人多能干细胞中腺病毒基因转移的优化
DOI：
10.1016/j.bbrc.2021.01.009
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Kosai K.
中科院分区：
文献类型：
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作者：
Mitsui K.;Takahashi T.;Ide K.;Matsuda E.;Kosai K.
人类多能干细胞（human pluripotent stem cells，human plu为了了解多能干细胞向功能细胞分化的过程，需要高效、安全地转移和表达外源基因。我们试图优化使用腺病毒载体将基因有效转移到多能干细胞中。对三种代表性的普遍活性启动子的活性的比较研究揭示，只有CA启动子允许在人多能干细胞中稳健的转基因表达。此外，我们还建立了一个方案，使我们能够有效地引入靶基因，并确保它们即使在少量细胞中也能表达。腺病毒载体感染单细胞悬浮培养中的多能干细胞产生了高的基因转移效率和低的细胞毒性，而不会失去多能干细胞的未分化状态。这种优化的系统将促进发育生物学研究和使用多能干细胞的再生医学。
Human pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells, have the potential to differentiate into a wide variety of cellsin vitroand 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.