E-GEOD-2585 - Transcription profiling of mouse thymic stromal cells from 6-12 week old wild type (C57BL/6) and Aire knock-outs (mixed background)
Submitted on 27 April 2005, released on 15 December 2007, last updated on 27 March 2012
Gene expression in different thymic stromal cells and subsets thereof was analyzed in 6-12 week old wild type (C57BL/6) and Aire knock-out (mixed background) mice. Thymic stromal cells were purified by sequential enzymatic digestion (collagenase, collagenase/dispase and trypsin) followed by gradient centrifugation and FACS sorting. Sort criteria were as follows: dendritic cells (CD11c+, F4/80 -), macrophages (F4/80+, CD11c-), cTECs (CD45–/lo, CDR1/Ly51+, Ep-CAM+) and mTECs (CD45–/lo, CDR1/Ly51–, Ep-CAM+). mTECs of wild-type and Aire knock-out mice were further subdivided according to CD80 expression levels.,For microarray analysis total RNA from thymic stromal cell samples of two independent experiments was pre-amplified and biotinylated by two rounds of cDNA synthesis and in vitro transcription. Fluorescence readings were evaluated by using Microarray Suite 5.0 software.
transcription profiling by array, cell type comparison, co-expression, individual genetic characteristics
Promiscuous gene expression in thymic epithelial cells is regulated at multiple levels. Jens Derbinski,Jana Gäbler,Benedikt Brors,Sascha Tierling,Sunitha Jonnakuty,Manfred Hergenhahn,Leena Peltonen,Jörn Walter,Bruno Kyewski. The Journal of experimental medicine :33-45 (2005)