Anti human cd45 conjugated microbeads

Surgically resected glioblastoma tissues were immediately digested with a papain-based dissociation system (Worthington, Lakewood, NJ). After digestion, leukocytes were depleted by using anti-human CD45-conjugated microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) and MACS LD columns (Miltenyi Biotec). The tumor cell population was filtered by excluding CD31-expressing cells. The single-cell quantitative RT–PCR (qRT–PCR) was performed at the Federal Institute of Technology (ETH) Zurich, Department of Biosystems Science and Engineering (D-BSSE) and Genomics Facility in Basel by using C1 Single-Cell Autoprep and BioMark HD instruments (Fludigim, San Francisco, CA). Cq values were converted to expression levels using the the equation Log₂Ex = C_{q LOD (Limit of Detection)}—Cq with a LOD C_q of 25 and data was mean-centered^{40 (link)}.

Anti-human CD45-conjugated microbeads were from Miltenyi Biotech (Bergisch Gladbach, Germany). Cell purification was performed according to the manufacturer’s protocol.