Adenovirus vector-mediated efficient transduction into human embryonic and induced pluripotent stem cells.
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.
Tashiro
Katsuhisa
K
National Institute of Biomedical Innovation, Osaka, Japan.
Kawabata
Kenji
K
Inamura
Mitsuru
M
Takayama
Kazuo
K
Furukawa
Norihisa
N
Sakurai
Fuminori
F
Katayama
Kazufumi
K
Hayakawa
Takao
T
Furue
Miho Kusuda
MK
Mizuguchi
Hiroyuki
H
D000256:Adenoviridae / Q000235:genetics*; D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D007763:Lac Operon; D011991:Receptors, Virus / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic
C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D017931:DNA Primers; D011991:Receptors, Virus