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Wild type and aire -/- murine meduallary thymic epithelial cells

ABSTRACT: Mice used were B6/129 F2's, 3-5 weeks of age, either wild type or with both copies of the autoimmune regulator gene (aire, GenBank #AF079536) disrupted. Thymi from five of these mice of both sexes were removed and pooled. After collagenase/dispase digestion, density gradient fractionation, and fluorescent antibody staining, cells with the phenotype CD45-, G8.8+, CDR1int and B7.1hi were FACS-sorted and total RNA was made from them. RNA was twice-amplified using a T7 polymerase-based method. Keywords: ordered

ORGANISM(S): Mus musculus

PROVIDER: GSE85 | GEO | 2002/09/19

SECONDARY ACCESSION(S): PRJNA85463

REPOSITORIES: GEO

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Transcription profiling of wild type and aire -/- murine meduallary thymic epithelial cells to study the immunological roles of aire protein in thymus.

Project description:Mice used were B6/129 F2's, 3-5 weeks of age, either wild type or with both copies of the autoimmune regulator gene (aire, GenBank #AF079536) disrupted. Thymi from five of these mice of both sexes were removed and pooled. After collagenase/dispase digestion, density gradient fractionation, and fluorescent antibody staining, cells with the phenotype CD45-, G8.8+, CDR1int and B7.1hi were FACS-sorted and total RNA was made from them. RNA was twice-amplified using a T7 polymerase-based method.

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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)

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Gene expression analysis of a human thymic cell line model of AIRE molecular function

Project description:Microarray analysis of AIRE regulated gene expression has been performed extensively in studies using the thymi of different mouse strains, but to date, has only been rarely performed using human model systems, and critically, not yet with a human thymic model. We have generated a Flp-In host cell line variant of the TEC 1A3 human thymic epithelial cell line and subsequently used site-specific recombination to generate a variant that stably expresses recombinant AIRE. We used microarray analysis to determine the transcription expression profiles of these two variants and determine the repertoire of AIRE regulated gene expression in a human thymic model system. The aim of the present study is to use microarray analysis to identify the repertoire of AIRE regulated genes within a human model system, and to validate this model as an appropriate human model system for the study of the molecular function of AIRE

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