I am really intrigued by this new approach and I am considering to adopt it. It seems very reliable and accurate. What are the main advantages you found, compared to other methodologies for gene expression analysis and cell characterisation? Any specific troubleshooting encountered? When using the method to characterise embryo stem cells, do you still proceed with the immunefluorescence analysis of EBs (as a" visual counterproof")? Did anyone use the method to characterise primary cells (derived from surgical waste)? And last, but not least: costs of the software?
http://www.lifetechnologies.com/it/en/home/life-science/stem-cell-research/taqman-hpsc-scorecard-panel.html?icid=PSM2-Scorecard-701
The main advantage of the TaqMan® hPSC Scorecard™ Assay is its ability to benchmark the gene expression of your ESC, iPSC and EBs against a reference set of well characterized ES and iPS lines. The assay was designed specifically to evaluate pluripotency and early germ layer specification, and was not designed for characterization of primary or terminally differentiated cells.
A common challenge that researchers experience when first using the TaqMan® hPSC Scorecard™ Panel is its high sensitivity to differentiation in the culture. For example, an 80% SSEA4 positive culture will be called as a "pass" for trilineage differentiation (ecto, meso, endo) and thus fail the test for the undifferentiated state, which requires both the presence of self-renewal factor expression and the absence of germ layer factor expression.
The hPSC Scorecard™ Analysis Software is available at no charge to the research community and can be accessed at http://apps.lifetechnologies.com/hPSCscorecard/home.htm From there, you can create a demo project to explore the various plots, reports and Excel exports of the data provided by the software. The demo project includes data for a one week time course of EB formation with H9 ESC.
A recent publication comparing the TaqMan® hPSC Scorecard™ Assay to the teratoma formation assay can be downloaded here: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0081622
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