Back to Search
Start Over
Adenovirus vector-mediated efficient transduction into human embryonic and induced pluripotent stem cells.
- Source :
-
Cellular reprogramming [Cell Reprogram] 2010 Oct; Vol. 12 (5), pp. 501-7. - Publication Year :
- 2010
-
Abstract
- We examined the transduction efficiency in human embryonic stem (ES) and induced pluripotent stem (iPS) cells using an adenovirus (Ad) vector. RT-PCR analysis revealed the expression of the coxsackievirus and adenovirus receptor, a receptor for Ad, in these cells. However, gene expression after the transduction with an Ad vector was observed only in the periphery of ES and iPS cell colonies, when human ES and iPS cells were passaged as small colonies. This suggests that the Ad vector could not enter inside the ES and iPS cell colonies by their tight connection. We thus attempted to transduce foreign genes into the dissociated form of human ES and iPS cells, which were passaged using Rho-associated kinase inhibitor. In this condition, transduction efficiency in human ES and iPS cells was markedly increased and transgene expression was observed even inside the colonies by using Ad vectors. Furthermore, Ad vector-mediated transduction did not alter the expression of undifferentiated markers such as Oct-3/4, Nanog, and SSEA-4. Our results indicate that Ad vectors are effective tools for transduction into human ES and iPS cells.
- Subjects :
- Base Sequence
Cell Differentiation
Cell Line
Coxsackie and Adenovirus Receptor-Like Membrane Protein
DNA Primers genetics
Embryonic Stem Cells cytology
Gene Expression
Genetic Vectors
Humans
Induced Pluripotent Stem Cells cytology
Lac Operon
Receptors, Virus genetics
Reverse Transcriptase Polymerase Chain Reaction
Adenoviridae genetics
Embryonic Stem Cells metabolism
Induced Pluripotent Stem Cells metabolism
Transduction, Genetic
Subjects
Details
- Language :
- English
- ISSN :
- 2152-4998
- Volume :
- 12
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Cellular reprogramming
- Publication Type :
- Academic Journal
- Accession number :
- 20726775
- Full Text :
- https://doi.org/10.1089/cell.2010.0023