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For induction of TGF-β 1 , spleen cells were cultured in serum-free medium X-Vivo-20 (BioWhittaker, Inc., Walkersville, MD, USA) in the presence of proteoglycan.
CV-1 cells were maintained in 10% fetal bovine serum (Bio-Whittaker, Walkersville, MD), 100 u/ml penicillin, 100 μg/ml streptomycin in DMEM.
Purified bovine CL-derived microvascular endothelial cell populations were generated commercially by Cambrex Biosciences (BioWhittaker, Walkersville, MD) as described in detail [ 35 ] . In the present study, CLENDO cells from frozen aliquots (passage 3) were seeded (5,000 cells/cm 2) in 6- or 24-well plates or 10 cm culture dishes.
Cell culture medium, nutrients and antibiotics were purchased from BioWhittaker (Walkersville, MD).
Cells were cultured in DMEM/F-12 medium (Sigma) supplemented with 20% (v/v) FCS (Biowhittaker Inc., Walkersville, MD) under 5% CO 2 in air at 37°C.