Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse thymic stromal cells from 6-12 week old wild type (C57BL/6) and Aire knock-outs (mixed background)


ABSTRACT: 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.

ORGANISM(S): Mus musculus

SUBMITTER: Benedikt Brors 

PROVIDER: E-GEOD-2585 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Promiscuous gene expression in thymic epithelial cells is regulated at multiple levels.

Derbinski Jens J   Gäbler Jana J   Brors Benedikt B   Tierling Sascha S   Jonnakuty Sunitha S   Hergenhahn Manfred M   Peltonen Leena L   Walter Jörn J   Kyewski Bruno B  

The Journal of experimental medicine 20050627 1


The role of central tolerance induction has recently been revised after the discovery of promiscuous expression of tissue-restricted self-antigens in the thymus. The extent of tissue representation afforded by this mechanism and its cellular and molecular regulation are barely defined. Here we show that medullary thymic epithelial cells (mTECs) are specialized to express a highly diverse set of genes representing essentially all tissues of the body. Most, but not all, of these genes are induced  ...[more]

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