Babco, Signet, Sternberger Monoclonals, Senetek, Covance

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Cell-Vive™ GMP CD137 (4-1BB) Activation Beads

422614

2.5 mL

1235 USD

FC, Cell Culture, Activ

422614

S18014C.Rec

Cell-Vive™ GMP CD137 (4-1BB) Activation Beads

2.5 mL

1235 USD

Human

CV_BEADS

Cell Vive™ GMP CD137 (4-1BB) Activation Beads are designed for the in vitro or ex vivo expansion of human T cells and NK cells. This product is a ready to use solution, and there is no need to dilute or reconstitute the product prior to use in cell culture.; Cell Vive™ GMP CD137 (4-1BB) Activation Beads are designed to assist in the activation and expansion of human T cells or NK cells, in tandem with other cell activation methods or alone. When used alone, this product is best utilized for re-stimulation.; This product has been optimized on human PBMCs, isolated human CD3+ T cells, and isolated CD56+ NK cells from whole blood. A suggested protocol for these cell types is below, but this protocol will require optimization depending on the intended use and cell source.; Suggested T cell expansion protocol in tandem with Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads (cat# 422605):;Prepare desired T cell complete media (TCCM) using sterile, filtered cell-culture grade reagents. Standard TCCM formulation: IMDM with 5% Cell-Vive™ T-NK Xeno-Free Serum Substitute, GMP (cat#: 420502) and 200 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925).;Prepare cells at a concentration of 1 x 10^6 per mL of TCCM.;Prepare the activation bead mixture of Cell-Vive™ GMP CD3/CD28 Human T Cell Activation Beads and Cell-Vive™ GMP 4-1BB/CD137 Activation Beads by mixing desired volume of beads in a 5 mL polypropylene tube with 1 mL of TCCM.When using PBMCs, ratios of CD3/CD28 beads and 4-1BB beads of 1:1, 1:3, and 1:4 have shown similar levels of fold expansion and final CD3+/CD8+ percentages. The optimal ratio of beads will depend on the cell source, media formulation, and cell culture scale.;If using a 1:1 bead-to-cell ratio and a 1:1 CD3/CD28-to-4-1BB bead ratio, mix 12.5 µL of each bead type for a final bead mixture volume of 25 µL.;Wash the beads by applying the tube with beads to a magnet (cat# 480019) for 1- 2 minutes. Decant supernatant and remove tube from the magnet.;Resuspend the beads in 1 mL of cell suspension and plate in a 12 well cell culture plate.;Allow cells to expand at 37°C and 5% CO2. At D4 and D7, count cells and re-seed at 0.5 x 10^6 cells per mL.;At D10 or D11, harvest cells for desired applications or re-stimulation.; Suggested NK cell expansion protocol in tandem with Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads (cat# 422611):;Prepare desired NK cell complete media (NKCCM) using sterile, filtered cell-culture grade reagents. A suggested NKCCM formulation is: IMDM with 5% human AB serum or serum substitute, 100 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925), and 15 ng/mL of Cell-Vive™ GMP Recombinant Human IL-15 (carrier-free) (cat#: 570314).;Prepare cells at a concentration of 1 x 10^6 per mL of TCCM.;Prepare the activation bead mixture of Cell-Vive™ GMP NKp46/CD2 Human NK Cell Activation Beads and Cell-Vive™ GMP 4-1BB/CD137 Activation Beads by mixing desired volume of beads in a 5 mL polypropylene tube with 1 mL of NKCCM.When using PBMCs, a ratio of NKp46/CD2 beads and 4-1BB beads of 1:1 and 1:3 have shown optimal levels of fold expansion. The optimal ratio of beads will depend on the cell source, media formulation, and cell culture scale.;If using a 1:1 bead-to-cell ratio and a 1:3 NKp46/CD2-to-4-1BB bead ratio, mix 6.25 µL of NKp46/CD2 beads and 18.75 µL of 4-1BB beads for a final bead mixture volume of 25 µL.;Wash the beads by applying the tube with beads to a magnet (cat# 480019) for 1-2 minutes. Decant supernatant and remove tube from the magnet.;Resuspend the beads in 1 mL of cell suspension and plate in a 12 well cell culture plate.;Allow cells to expand at 37°C and 5% CO2. At D6, D9, and D12, count cells and re-seed at 1.0 x 10^6 cells per mL.;At D14, harvest cells for desired applications or re-stimulation.; Suggested re-stimulation protocol for T cells or NK cells:;Prepare desired cell culture media using sterile, filtered cell-culture grade reagents.A suggested NK cell complete media (NKCCM) formulation is: IMDM with 5% human AB serum or serum substitute, 100 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925), and 15 ng/mL of Cell-Vive™ GMP Recombinant Human IL-15 (carrier-free) (cat#: 570314).;A suggested TCCM formulation is: IMDM with 5% Cell-Vive™ T-NK Xeno-Free Serum Substitute, GMP (cat#: 420502) and 200 IU/mL of Cell-Vive™ GMP Recombinant Human IL-2 (E. coli expressed, carrier-free) (cat#: 791925).;Prepare cells at a concentration of 1 x 106 per mL of media.;Prepare the Cell-Vive™ GMP 4-1BB/CD137 Activation Beads for wash by mixing 25 µL. of beads in a 5 mL polypropylene tube with 1 mL of media.This volume will provide a 1:1 bead-to-cell ratio for re-stimulation. The optimal bead-to-cell ratio should be determined experimentally for each cell culture protocol and cell type.;Wash the beads by applying the tube with beads to a magnet for 1-2 minutes. Decant supernatant and remove tube from the magnet.;Resuspend the beads in 1 mL of cell suspension and plate in a 12 well cell culture plate.;Allow cells to expand at 37°C and 5% CO2.For T cells, count cells on D4 and D7. Re-seed at 0.5 x 10^6 cells per mL.;For NK cells, count cells on D6, D9, and D12. Re-seed at 1.0 x 10^6 cells per mL.;At desired endpoint, harvest cells for desired applications and analysis.;