Human monocytic cell line THP1 (authenticated and tested mycoplasma free), obtained from the Department of Hematology, Erasmus Medical Center, was maintained in culture in RPMI-1640 (Lonza, Breda, The Netherlands) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. THP1 cells were differentiated into macrophage-like cells by PMA (Sigma, Stockholm, Sweden) at a concentration of 100 ng/ml for 48 h. Recombinant human CECR1 protein (rhCECR1) was added to the macrophage cultures at a concentration gradient of 0, 12.5, 25, 50, 100, 200 nm for 96 h.
HUVECs (Lonza) and HUVECs transfected with a lentiviral vector encoding GFP were cultured in EGM-2 endothelial medium (Lonza) with 1% penicillin/streptomycin.
Human brain vascular pericytes with and without lentiviral transfection of a vector encoding for dsRed, and GBM cell lines U87 and U251, purchased from ATCC, were maintained in DMEM (Lonza) with 10% fetal bovine serum and 1% penicillin/streptomycin. Pericytes were treated with recombinant human PDGFBs (Sigma) at different concentrations according to the specifications of the different assays. All cultured cells were checked to be negative for mycoplasma contamination. HUVECs and pericytes were authenticated by supplier and were tested mycoplasma free.
HUVECs (Lonza) and HUVECs transfected with a lentiviral vector encoding GFP were cultured in EGM-2 endothelial medium (Lonza) with 1% penicillin/streptomycin.
Human brain vascular pericytes with and without lentiviral transfection of a vector encoding for dsRed, and GBM cell lines U87 and U251, purchased from ATCC, were maintained in DMEM (Lonza) with 10% fetal bovine serum and 1% penicillin/streptomycin. Pericytes were treated with recombinant human PDGFBs (Sigma) at different concentrations according to the specifications of the different assays. All cultured cells were checked to be negative for mycoplasma contamination. HUVECs and pericytes were authenticated by supplier and were tested mycoplasma free.