Affiliations: Cellular Neurobiology Research Branch, Intramural
Research Program, National Institute on Drug Abuse, National Institutes of
Health, Department of Health and Human Services, Baltimore, MD, USA
Abstract: Purpose: human embryonic stem cells (hESCs) which express a reporter
gene consistently during all phases of differentiation would be valuable for
basic research on cell transplantation. In this study, we describe
karyotypically-abnormal variant hESCs, BGO1V2-EFG, which express hrGFP driven
by the EF1 promoter. Methods: BGO1V2-EFG cells were analyzed by using
immunocytochemistry, single cell-based confocal image, and in vitro
differentiation, including dopaminergic differentiation. Results: Undifferentiated BGO1V2-EFG cells expressed pluripotent ESC
markers and retained the ability to differentiate into cell types of all three
germ layers. BGO1V2-EFG cells maintained stable and robust hrGFP expression in
vitro in the undifferentiated state and during differentiation. The EF1
promoter retained activity during dopaminergic differentiation, as 76% of
tyrosine hydroxlase (TH)-positive cells co-expressed hrGFP by confocal
analysis. Treated with sodium butyrate (0.02 mM to 2.0 mM), an inhibitor of
histone deacetylase (HDAC), during differentiation did not affect hrGFP
expression, although TH expression was reduced by higher concentrations of
sodium butyrate. Conclusion: BGO1V2-EFG cells maintain stable and robust hrGFP
expression in the undifferentiated state and during neural differentiation.
Especially, the EF1 promoter was effective in driving hrGFP expression during
dopaminergic differentiation. BGO1V2-EFG cells may be useful for
transplantation studies in Parkinson disease animal models.
