Leukapheresis products from anonymous healthy human donors were purchased from STEMCELL Technologies, Inc. and isolated using an EasySep human T-cell isolation kit (Cat #17951). Isolated CD3+ T cells were activated at 1 × 106 cells/ml in a 1:1 ratio with CD3/CD28 magnetic dynabeads (CTS, Thermo Fisher Scientific), 100 U/ml IL-7 and 10 U/ml IL-15 (R&D Systems) for 48 h in complete XVivo15 medium (Lonza) (5% FBS, 50 μM 2-mercaptoethanol, 10 mM N-acetyl l -cysteine). Following the 48-h activation period, CD3+ T cells were debeaded with an EasySep magnet (STEMCELL Technologies, Inc.) prior to resuspension and electroporation at 0.5–1 × 106 cells/ml in P3 buffer (Lonza). Following electroporations with RNPs and HDR templates, fresh medium and cytokines were added every 2–3 days.
Cd3 cd28 magnetic dynabeads cts
Manufactured by Thermo Fisher Scientific
The CD3/CD28 magnetic dynabeads (CTS) are laboratory equipment used for the activation and expansion of T cells. They consist of superparamagnetic beads coated with antibodies against the CD3 and CD28 surface receptors on T cells. These beads can be used to stimulate T cell proliferation and differentiation in cell culture applications.
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Lab products found in correlation
2 protocols using cd3 cd28 magnetic dynabeads cts
1
T-cell Activation and Genome Editing
2
Isolation and Activation of CD3+ T Cells
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Leukapheresis products from anonymous healthy human donors were purchased from STEMCELL Technologies, Inc. and isolated using an EasySep human T cell isolation kit (Cat #17951). Isolated CD3+ T cells were activated at 1x10 6 cells mL -1 in a 1:1 ratio with CD3/CD28 magnetic dynabeads (CTS, Thermo Fisher Scientific), 100 U mL -1 of IL-7, and 10 U mL -1 IL-15 (R&D Systems) for 48 hours in complete XVivo15 medium (Lonza) (5% fetal bovine serum, 50 µM 2-mercaptoethanol, 10 mM N-alcetyl L-cysteine).
Following the 48-hour activation period, CD3+ T cells were debeaded with an EasySep magnet (STEMCELL) prior to resuspension and electroporation at 0.5-1x10 6 cell mL -1 in P3 buffer (Lonza). Following electroporations with RNPs and HDRTs, fresh medium and cytokines were added every 2-3 days.
Following the 48-hour activation period, CD3+ T cells were debeaded with an EasySep magnet (STEMCELL) prior to resuspension and electroporation at 0.5-1x10 6 cell mL -1 in P3 buffer (Lonza). Following electroporations with RNPs and HDRTs, fresh medium and cytokines were added every 2-3 days.
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