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Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed

Product Size Amount
Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed 2.5 mg 2.5 × 107 beads
10 mg 1 × 108 beads

背景(Background)

Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed are uniform 5.5 μm of magnetic beads coated with an optimized mixture of monoclonal antibodies against the CD3 and CD28. The CD3 antibody is specific for the epsilon chain of human CD3, which is considered to be a subunit of the TCR complex. The CD28 antibody is specific for the human CD28 co-stimulatory molecule, which is the receptor for CD80 (B7-1) and CD86 (B7-2). Both antibodies are mouse anti-human IgGs coupled to the same bead, mimicking in vivo stimulation by APCs.

应用(Application)

Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed is intended for the in vitro stimulation and expansion of purified T cell populations of, for example, CD3+T cells, CD4+ T cells, CD8+ T cells or human PBMC.

This is for research use and ex vivo cell culture processing only, and is not intended for human in vivo applications.

无菌(Sterility)

The sterility testing was performed by membrane filtration method.

内毒素(Endotoxin)

Less than 2EU/mg by the LAL method.

重构方法(Reconstitution)

See Certificate of Analysis (CoA) for detailed instruction.

存储(Storage)

This product is stable after storage at: -20˚C for 24 months in lyophilized state; 2˚C to 8˚C for 24 months under sterile conditions after reconstitution.

Please avoid freezing after reconstitution. Immediate use after reconstitution is highly recommended.

注意事项(Important Note)

This product is for research use only and not intended for therapeutic or in vivo diagnostic use.

 

Data

Activation human T cells analyzed by FACS

Fig.1. Activation of the purified human T Cells. The purified human T cells were activated using Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed (Cat. No. MBS-C001) at a ratio of 1:1 beads-to-cells for 24 hours with RPMI1640 supplemented with 10% of FBS. The negative control experiment was performed with adding the Negative Control Beads coupled HSA. Cells were fluorescently stained using PE labeled anti-human CD25 antibody and labeled FITC anti-human CD69 antibody and analyzed by flow cytometry.

Protocol

Activation of the human T Cells expansion

Fig.2. Purified human T Cells expansion. The purified human T cells were stimulated using Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed (Cat. No. MBS-C001) at a ratio of 1:1 beads-to-cells. Cells were expanded in T cell culture medium supplemented with 4ng/mL of rhIL-2 Protein (Acrobiosystems, Cat. No. IL2-H4113). Activated Cells were expanded for up to 13 days (A) with high cell viability (B).

Protocol

Low level T-cell exhaustion

Fig.3. PD-1 expression of the activated human T Cells. The purified human T cells were stimulated using Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed (Cat. No. MBS-C001) at a ratio of 1:1 beads-to-cells. Cells were expanded in T cell culture medium supplemented with 4ng/mL of rhIL-2 Protein (Acrobiosystems, Cat. No. IL2-H4113). Activated T cells were expanded for up to 10 days with low PD-1 expression.

Protocol

Activation in different medium

Fig.4.Activation of the purified human T Cells with IL-2 in different medium.The purified human T cells were stimulated using Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed (Cat. No. MBS-C001) at a ratio of 1:1 beads-to-cells in three commercial medium, Human rh IL-2 (Cat. No. IL2-H5215, 4ng/mL) was added. The expansion ability of T cells varied greatly in the culture medium from different manufacturers.

Activation comparison with competitor

Fig.5.Activation of the purified human T Cells. The purified human T cells were activated using Human T cell Activation/Expansion CD3/CD28 Beads, premium grade DMF Filed (ACRO,Cat. No. MBS-C001) and competitor respectively for 24 hours. Cells were fluorescently stained using PE labeled anti-human CD25 antibody and labeled FITC anti-human CD69 antibody and analyzed by flow cytometry.

Protocol

Purified human T Cells expansion

Fig.6.Purified human T Cells expansion. The purified human T cells were stimulated using Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) at a ratio of 1:1 beads-to-cells. Cells were expanded in T cell culture medium supplemented with 4ng/mL of rhIL-2 Protein (Acrobiosystems, Cat. No. IL2-H4113). Activated Cells used Acro were expanded more for 14 days with higher cell expansion fold and viability compared with Competitor. (Data from different donors are similar, but only one donor are presented)

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