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Human PBMCs

Manufactured by AllCells
Sourced in United States

Human PBMCs are a type of peripheral blood mononuclear cells (PBMCs) isolated from human blood. PBMCs are a heterogeneous population of cells that include lymphocytes, monocytes, and dendritic cells. These cells can be used for various in vitro applications, such as cell culture, immunological studies, and drug testing.

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5 protocols using Human PBMCs

Human tumor cell lines U937, THP-1, Raji and SK-BR-3 were obtained from the American Type Culture Collection and cultured as recommended by the vendor. Human PBMCs, CD14+ monocytes, neutrophils and dendritic cells were purchased from AllCells.
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Frozen human PBMCs were obtained from AllCells. The T cell culture medium (TCM) was composed of AIM-V medium with 5% (v/v) human AB serum, 10 mM HEPES, 1% (v/v) GlutaMax-100×, 12.25 mM N-acetylcysteine (NAC), 100 U/mL penicillin, and 100 μg/mL streptomycin. The cell culture was supplemented with 10 ng/mL human IL-2. After the activation by Dynabeads Human T-Expander CD3/CD28 (Invitrogen) at a bead:PBMC ratio of 3:1 for 48 h, PBMCs were transduced with retroviral vectors using the method previously described. Transduced T cells were expanded for 2 weeks in TCM, during which, time-culture medium was replenished every 2 days, and T cell density was maintained between 0.5 and 1 × 106 cells/mL.
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NCI-N87, HCC1954, and SKBR3 cells were obtained from the Shanghai Cell Line Bank. RPMI1640 medium (61870036), Freestyle 293 medium, Dynabeads CD4 Positive Isolation Kit (11331D), TMB substrate, and fetal calf serum (FCS) were obtained from Thermo Fisher Scientific. PE-labeled goat anti-human IgG (409304), streptavidin-PE (405203), anti-human-CD3 (clone HIT3a), and anti-human-CD28 (clone CD28.2) were obtained from Biolegend. Trastuzumab and humanized anti-PD-L1 scFV were prepared in house. Cell Counting Kit-8 (CCK8) was obtained from Dojindo Laboratories. Human antibody germline sequence and primers were synthesized by Sangon Biotech. IL-4 and GM-CSF were obtained from R&D Systems, and all other recombinant proteins were products of Sino Biological. SYPRO Orange was obtained from Sigma-Aldrich. Human IFN-γ ELISA Kit (EHC102g) was obtained from Neobioscience. Cytotoxicity detection kit (LDH) was from Roche Life Science. Matrigel was obtained from BD Biosciences. Polyethylenimine (PEI) was obtained from Polysciences. Protein A probes were obtained from Pall Corporation. Protein A-Sepharose Column was obtained from BioVision. HRP-conjugated goat anti-human IgG (H + L) was obtained from Jackson ImmunoResearch. Monocyte purification kit was obtained from Miltenyi Biotec. Ficoll gradient was obtained from GE healthcare. Human PBMCs were obtained from Allcells.
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Human PBMCs (AllCells, Alameda, CA, USA) were cultured in AIM V® Medium CTS (Gibco, USA). K562 cells were cultured in RPMI 1640 medium (Gibco, USA) with 10% fetal bovine serum and 1% penicillin–streptomycin (Gibco, USA).
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Frozen, de-identified human PBMCs (Allcells, Alameda, CA) were thawed and stimulated with 6 IU IL-2 (R&D Systems, Minneapolis, MN) and 10 μg/ml concanavalin A (ConA, MilliporeSigma, Billerica, MA) in RPMI. Six days post-thaw, CD4+ T cells were positively isolated using an EasySep Human CD4 positive selection kit in a RoboSep robot (#18052RF, Stemcell Technologies, Cambridge, MA). 2D10 Jurkat cells (38 (link)) were also grown in RPMI. L22 selectively labeled with 5/6-carboxyfluorescein was added at indicated concentrations at indicated time points prior to flow cytometry analysis as described above.
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