DNT expansions were performed as previously described.11 (link) Briefly, CD4+ and CD8+ cell-depleted healthy donor–derived PBMCs were cultured on anti-CD3 antibody-coated plates (OKT3; BioLegend) for 3 days in AIM-V (Thermo Fisher Scientific) with 250 IU/mL of interleukin-2 (IL-2) (Proleukin; Novartis Pharmaceuticals); soluble anti-CD3 antibody and IL-2 were added to the cultures every 2 to 4 days. The purity of DNTs was assessed by staining cells with fluorochrome-conjugated anti-human CD3, -CD4, -CD8, and -CD56 antibodies and flow cytometry analysis. DNTs were used between days 10 and 20 of culture. For venetoclax treatment, 25 to 400 nM venetoclax (ABT-199; TargetMol) was added to DNT culture 4 hours to 3 days before use. DNTs were washed twice with phosphate-buffered saline and counted before use for each experiment.
Venetoclax
Manufactured by Targetmol
Venetoclax is a small-molecule inhibitor of the anti-apoptotic protein BCL-2, which is involved in regulating cell survival and death. It is used in the research and development of cancer therapeutics.
Automatically generated - may contain errors
2 protocols using venetoclax
1
Expanding and Treating DNT Cells
2
Venetoclax and Fedratinib Combination Assay
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Venetoclax (1 mL*10 mM in DMSO) and powdered compound were purchased from TargetMol. Fedratinib (1 mL*10 mM in DMSO) was purchased from Selleckchem and powdered was purchased from MedChemExpress. All concentrations for in vitro studies were made at 2x or 4x for combination studies and diluted in RPMI-10 for plating assays. Cells were treated with a range of doses ranging from 1nM to 10uM.
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