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IN Cell User's Day 2010 講演要旨

High Content Analysis for stem cell and GPCR screening

Andrew J. Ball Ph. D
High Content Analysis Program Leader
Millipore Corporation
St. Charles, Missouri, USA

The need for more biologically relevant applications that improve decision-making power for drug discovery has never been greater. Millipore's innovative research and development programs continue to focus on meeting this need. Using a GE IN Cell Analyzer High Content Analysis (HCA) platform as a primary development tool, a goal has been to develop extensively validated cellular imaging assays for multiple stages in the drug discovery process, from primary screening thru in vitro toxicity assessment. Case studies will be presented illustrating the latest High Content Analysis applications for stem cell profiling and for GPCR drug discovery, combining cellular imaging and stem cell-derived model systems. Particular attention will focus on a novel HCA assay for extracellular signal-regulated kinase (ERK). ERK signaling controls cell growth, differentiation and survival. ERK is activated by dual phosphorylation on tyrosine and threonine residues (pERK). This pathway is often upregulated in tumors and is a target for cancer therapy. Recent evidence suggests that pERK may also represent a generic readout for GPCR activation. Using the IN Cell Analyzer, we have developed an immunofluorescence-based cellular imaging assay for ERK activation, in order to quantitate the activity of agents modulating pERK. Experiments demonstrated the application of this assay to screen for GPCR activation in response to receptor activation in control cells and engineered GPCR cell lines. We then demonstrate that this assay is a useful tool for profiling GPCR activation in a panel of human embryonic stem cell lines. In conclusion, applications of HCA for drug discovery continue to grow, and the IN Cell Analyzer represents an exciting technology for new assay development.


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