293T (HEK 293 T/17), Raji, Daudi, and THP-1 cell lines were obtained from the American Type Culture Collection. Cell lines were maintained in DMEM (Invitrogen, Grand Island, NY) supplemented with 10% fetal calf serum (FCS) and 2 mM GlutaMAX™ (Invitrogen) at 37 °C and 5% CO2. T cells generated from peripheral blood mononuclear cells (PBMC) were cultured in 45% RPMI 1640, 45% Click’s media (Invitrogen) supplemented with 10% fetal bovine serum (FBS), 2 mM GlutaMAX (T-cell media, TCM), and 100 U/ml IL-2 (Miltenyi Biotec, Bergisch Gladbach, Germany), unless otherwise noted. Clinical-grade rimiducid (5 mg/ml in 25% Kolliphor HS15®) was diluted in ethanol to a 100 mM working solution for in vitro assays, or 0.9% saline for animal studies.
Click s media
Manufactured by Thermo Fisher Scientific
Sourced in Germany
Click's media is a laboratory product designed to support cell culture and cell growth. It provides essential nutrients and growth factors to maintain and propagate cells in a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 2 (il 2), by Miltenyi Biotec (2 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Miltenyi Biotec (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Thp 1, by American Type Culture Collection (1 mentions)
Daudi, by American Type Culture Collection (1 mentions)
Anti cd3 and anti cd28 antibodies, by Miltenyi Biotec (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Genejuice, by Merck Group (1 mentions)
2 protocols using click s media
1
Cell Line Maintenance and T-Cell Culture
2
Transduction of Activated T Cells
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Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats obtained through the Gulf Coast Regional Blood Center, and they were cultured in 45% RPMI 1640 and 45% Click’s media (Invitrogen, Grand Island, NY), supplemented with 10% fetal bovine serum (FBS) and 100 U/ml IL-2 (Miltenyi Biotec). Buffy coats were tested negative for infectious viral pathogens. Retrovirus was produced by transient transfection of HEK293T cells using GeneJuice (EMD Millipore, Billerca, MA) with MoMLV gag-pol (PegPam3-e plasmid), RD114 envelope (RDF plasmid), and the pSFG retroviral vector encoding the transgenes. Supernatant was collected after 48–72 h to transduce T cells that were activated by stimulation with 0.5 μg/mL each of anti-CD3 and anti-CD28 antibodies (Miltenyi Biotec) in the presence of 100 U/mL IL-2. The spinfection technique was performed to transduce T cells with RetroNectin coating and expanded for 10–14 days post-transduction unless otherwise stated. For transductions with multiple vectors, the protocol was identical to the above, except the wells were coated with equal amounts of each retroviral supernatant.
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