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Recombinant AAV Transduction of hESCs.

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posted on 17.11.2011, 00:23 by Matthew L. Hirsch, B. Matthew Fagan, Raluca Dumitru, Jacquelyn J. Bower, Swati Yadav, Matthew H. Porteus, Larysa H. Pevny, R. Jude Samulski

The indicated AAV serotypes were packaged with a self-complementary CMV-egfp genome and used to infect human embryonic stem cells (hESCs) at 100,000 viral genomes per cell. 24 h post-infection (post-infection) cells were harvested and GFP+ cells were quantitated by flow cytometry (A.). Treated cells were also analyzed for intracellular transgene copy number normalized to the lamin B gene (B). Cell viability was also measured under the indicated conditions via dye exclusion (C) and significant decreases (p-value<0.05) were noted in all cases (compared to no virus) except with AAV4 treatment (p-value = 0.44). The results are averaged from at least 6 replicates for each treatment group and the standard deviation is depicted.

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