Characterizing ccRCC Tumor Cell Subsets

Six ccRCC specimens were obtained from the University Health Network (UHN) Program in Biospecimen Sciences from consenting patients in accordance with the policies of the UHN Research Ethics Board (REB ID# 09–0828). Flow cytometry was performed as previously described [27 (link)]. Briefly, primary ccRCC specimens were disaggregated into single cell suspensions using collagenase treatment. The specimens were stained using CD45-APC-Cy7 (1:200), CD31-PE-Cy7 (1:200), CD34-PerCP-Cy5.5 (1:50) (all BioLegend) and 1.2 mg/ml TE7-biotin (in-house production from hybridoma obtained from ATCC) followed by Streptavidin-eFluor450 (1:400; eBioscience). These cells were then stained as described in Gedye et al. with an antibody panel that included other stem/progenitor and immune markers [27 (link)]. Data collection was performed on a Becton-Dickinson LSR II flow cytometer.