CD47+ Jurkat cell line BioVector NTCC中国质粒载体菌种细胞基因保藏中心

CD47+ Jurkat cell line BioVector NTCC中国质粒载体菌种细胞基因保藏中心 Source name Jurkat cell line Organism Homo sapiens Characteristics cell type: CD47+ Jurkat cells genotype/variation: CD47+ JK exosome Treatment protocol No treatments were performed. Growth protocol Jurkat T cells and the corresponding CD47-deficient Jurkat mutant JinB8 were maintained at 2.0 x 105 cells per ml in RPMI 1640 medium (Life technologies) supplemented with 10% human serum AB, glutamine, penicillin, and streptomycin. Jurkat cells were maintained for a maximum of 4 weeks for experiments. Before exosome isolation, the cells were cultured using low human AB serum in their respective media for at least one passage. The T cells were cultured in 75 cm3 flasks (Corning) and conditioned medium from ~40x106 cells (~100 ml) was collected. The conditioned medium was further centrifuged for 10 min at 1530×g to remove cell debris. The supernatants were transferred to new 50 ml tubes. The supernatants were concentrated using centrifugal filter device YM-10 (Centricon). The final volume of supernatant was ~1 ml. The concentrated supernatants (~250 μl) from the respective cells were used to purify exosomes. The exosomes were isolated using the exoQuick kit (SBI System Biosciences) according to the manufactures instructions. The supernatant was completely removed via centrifugation 2 times for 30 min at 1500×g. The white exosome pellet was diluted into 1 ml of ultra-pure water (~10×106 cells). HUVEC were incubated with Jurkat and JinB8 T cell-derived exosomes using EGM2 media for 6 hours.