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Charles' presentation

High-Throughput Screening Assay for the Identification of Compounds Regulating Self-Renewal and Differentiation in Human Embryonic Stem Cells Sabrina C. Desbordes1, 2, 6, , , Dimitris G. Placantonakis2, 5, Anthony Ciro3, Nicholas D. Socci4, Gabsang Lee1, 2, Hakim Djaballah3 and Lorenz Studer1, 2,

In this publication Desbordes et. al. develop a high throughput technique for identification of compounds (small molecules) for thier ability to promote short-term self-renewal or drive early differentiation. First the cells needed to be conditioned to a sensitized state thus making them amenable to eather fate, self renewal or diffentiation, upon treatment with the compounds. To acheive this cells were seeded in a matrigel coated 384-well plate with FGF2 supplimentation until day 2 at which time the FGF2 supplimentaion was withdrawn. Oct4 expression was used as the measure of self-renewal. For the High control contued treatment with FGF2 for seven days and for the low control treatment with BMP4 was used to induce differntiation. So, at day 2 the cells were treated with either a compound or the high or low control until day 7. On day 7 the cells were stained with Oct4 antibody using Multidrop 384 and Flexdrop IV liquid handlers. The confocal image aquisition was carried out using the InCell Analizer 3000.