High-throughput screening for modulators of stem cell differentiation.
Realizing the potential of stem cell biology requires the modulation of self-renewal and differentiation, both of which are incompletely understood. This chapter describes methods for the design, development, and implementation of cell-based screens of small molecules, genes and expressed proteins for modulation of stem and progenitor cell fate. These include the engineering of embryonic and other stem cells with gene promoter-reporter protein constructs and their application in automated screening. We discuss considerations of promoter reporter selection, assay development and implementation, and image acquisition, analysis, and data handling. Such black-box screens are useful for the identification of probes of developmental processes and should provide tools that will identify druggable targets for biochemical assays.
Bushway
Paul J
PJ
Burnham Institute for Medical Research, La Jolla, CA, USA.
Mercola
Mark
M
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D004622:Embryo, Mammalian / Q000473:pathology; D017930:Genes, Reporter; D016678:Genome; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D013234:Stem Cells / Q000166:cytology*