期刊
STEM CELLS
卷 25, 期 11, 页码 2928-2935出版社
ALPHAMED PRESS
DOI: 10.1634/stemcells.2007-0468
关键词
sox2 transcription factor; Akt1; neural progenitor cells; soft lithography
资金
- NIGMS NIH HHS [T32 GM067545] Funding Source: Medline
- NINDS NIH HHS [NS048248] Funding Source: Medline
We describe a microarray-based approach for the high-throughput screening of gene function in stem cells and demonstrate the potential of this method by growing and isolating clonal populations of both adult and embryonic neural stem cells. Clonal microarrays are constructed by seeding a population of cells at clonal density on micropatterned surfaces generated using soft lithographic microfabrication techniques. Clones of interest can be isolated after assaying in parallel for various cellular processes and functions, including proliferation, signal transduction, and differentiation. We demonstrate the compatibility of the technique with both gain- and loss-of-function studies using cell populations infected with cDNA libraries or DNA constructs that induce RNA interference. The infection of cells with a library prior to seeding and the compact but isolated growth of clonal cell populations will facilitate the screening of large libraries in a wide variety of mammalian cells, including those that are difficult to transfect by conventional methods.
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