Expression data from thymic endothelial cells after damage

The thymus is extremely sensitive to damage but also has a remarkable ability to repair itself. However, the mechanisms underlying this endogenous regeneration remain poorly understood and this capacity diminishes considerably with age. To identify alternate regeneration pathways in the thymus, we performed an unbiased transcriptome analysis of the non-hematopoietic (CD45-) stromal cell compartment of the thymus, which is less sensitive to thymic damage compared to the CD45+ hematopoietic compartment. Recent work has suggested that ECs can contribute towards thymic regeneration via their production of so-called angiocrine factors. Using a technique to constitutively activate the Akt pathway in ECs using the pro-survival adenoviral gene E4ORF1, ECs can be propagated and expanded ex vivo (exEC) while maintaining their phenotype and vascular tube formation capacity. We compared the gene expression of freshly isolated thymic ECs and exEC-derived from the thymus. Thymic non-hematopoietic stromal cells were isolated using CD45 MACS cell depletion. Endothelial cells were then FACS purified (CD45- EpCAM- VE-Cad+). exEC were generated by lentiviral transduction with the E4ORF1 gene. Microarray analysis was performed on an Affymetrix MOE 430 A 2.0 platform in triplicate for freshly isolated ECs from untreated mice, and for exECs transduced with the pro-survival adenoviral gene E4ORF1.