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Online Data Supplements for Identification of a Novel Role of T cells in Postnatal Vasculogenesis : Characterization of Endothelial Progenitor Cell Colonies This supporting material contains: Supplemental Materials and Methods Supplemental figure 1~6 and with figure legends Supplemental Table 1 Supplemental Materials and Methods Cells All study protocols in this study were approved by the Institutional Review Board of Seoul National University Hospital and the Aeromedical Center, ROKAF (Republic of Korea Air Force). Peripheral blood (50~150 mL) was obtained from donors with informed consent. The mononuclear cells were isolated from peripheral blood of donors by density gradient centrifugation with Histopaque-1077 (Sigma) according to manufacturer’s instructions. Cell Culture As described in our previous reports,1,2 isolated mononuclear cells were suspended by EGM-2 MV BulletKit system (Clonetics) consisting of endothelial basal medium, 5% fetal bovine serum, hEGF, VEGF, hFGF, IGF-1, ascorbic acid, and heparin. 1 x 107 mononuclear cells per well were seeded on 2% gelatin or fibronectin coated six-well plates and incubated in a 5% CO2 incubator at 37°C. Under daily observation, first media change was performed at days 4~7 after plating. Thereafter, media were changed every 3 days. Each cluster or spindle-shaped cells were followed-up every day. Adherent cells staining positive for both FITC–ulex-lectin (Sigma) and DiI-acLDL (Molecular Probes) were considered as EPCs. Magnetic Activated Cell Sorting (MACS) Experimental scheme using MACS is as follows (supplemental Fig. 1): Untouched CD3+ T cells were enriched to high purity (98%) from PBMNCs by magnetic separation using Pan T Cell Isolation Kit II (MACS; Miltenyi Biotec, Bergisch Gladbach, Germany) which excluded cells positive for CD14, CD16, CD19, CD36, CD56, CD123 and Glycophorin A. These CD3+ T cells were then separated in a second magnetic separation step into CD3+CD31+ (Double Positive ; DP) and