The cell lines MDA-MB-231/GFP (Cell Biolabs) and NIH 3T3 (ATCC) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% penicillin-streptomycin. The NK-92 cells (ATCC) were cultured in Alpha minimum essential medium (ATCC, VA, USA) supplemented with 0.2 mM inositol (Sigma), 0.1 mM 2-mercaptoethanol (Sigma), 0.02 mM folic acid (Sigma), 100–200 U/ml recombinant interleukin-2 (PeproTech, NJ, USA), 12.5% horse serum, 12.5% FBS, and 1% penicillin-streptomycin. K562 cells (ATCC) were cultured in Iscove’s modified Dulbecco’s medium, supplemented with 10% (v/v) FBS and 1% penicillin-streptomycin. All cells were grown in a humidified atmosphere of 5% (v/v) CO2 at 37°C. Adherent cells (MDA-MB-231 and NIH 3T3) at 80% confluence in a 60 mm × 15 mm Petri dish were harvested by trypsin digestion and dispensed into a cell suspension with various concentrations according to the experimental requirement. Suspended cells (NK-92 and K562) were centrifuged and washed by cell-free media or PBS and finally adjusted into known cell centration.
Recombinant interleukin 2
Manufactured by Thermo Fisher Scientific
Sourced in United States, Japan
Recombinant interleukin-2 is a cytokine protein produced through recombinant DNA technology. It functions as a signaling molecule, playing a role in the activation and proliferation of T cells and natural killer cells in the immune system.
Automatically generated - may contain errors
Lab products found in correlation
Inositol, by Merck Group (1 mentions)
Nk 92 cells, by American Type Culture Collection (1 mentions)
Mda mb 231 gfp, by Cell Biolabs (1 mentions)
K562 cell, by American Type Culture Collection (1 mentions)
Nih3t3, by American Type Culture Collection (1 mentions)
Folic acid, by Merck Group (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
2 protocols using recombinant interleukin 2
1
Cell Culture Protocols for Cancer and Immune Cell Lines
2
Engineered CAR-T Cell Generation
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The constructs of CAR20, the expansion, culture and the verification of transduction efficiency of CART-20 cells were primarily performed as described previously in our clinical trial.15 (link) Briefly, peripheral blood mononuclear cells were activated with immobilized CD3 antibodies (Takara, Toyko, Japan) and recombinant interleukin-2 (500 U ml−1; PeproTech, Rocky Hill, NJ, USA). Lentivirus-mediated CAR transduction was performed on day 3 of the cell culture. After transduction, T-cell lines were expanded in the presence of interleukin-2 (500 U ml−1). The composition and purity were assessed by fluorescence-activated cell sorting, and the cells were harvested beginning on days 10–12.
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