ImmunoCult™ Human CD3/CD28 T Cell Activator
Human T cell activation and expansion reagent
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Overview
This product’s gentle activation stimulus ensures a high viability of activated T cells, which can be further expanded in ImmunoCult™-XF T Cell Expansion Medium (Catalog #10981) or other media for culturing human T cells. 1. Antibody complexes bind to and cross-link CD3 and CD28 cell surface ligands, thereby providing the required signals for T cell activation. ImmunoCult™ Human CD3/CD28 T Cell Activator can be used on the Seahorse XF Analyzer to measure T cell activation response and is also available as part of the Agilent Seahorse XF Hu T Cell Activation Assay Kit.
This product is designed for cell therapy research applications, but may be qualified for use as an ancillary material (AM) following the framework outlined in USP<1043>. STEMCELL can work with you to qualify this reagent as an AM under an approved investigational new drug (IND) or clinical trial application (CTA). Learn more about how we can support your regulatory needs here.
• Anti-human CD28 monospecific antibody complex
Data Figures

Figure 1. Activated Morphology of Human T Cells Stimulated With ImmunoCult™ Human CD3/CD28 T Cell Activator
Image of human T cells isolated using the EasySep™ Human T Cell Isolation Kit (Catalog #17951), stimulated with ImmunoCult™ Human CD3/CD28 T Cell Activator, and cultured in ImmunoCult™-XF T Cell Expansion Medium (Catalog # 10981).

Figure 2. Activation of EasySep™-isolated Human T Cells stimulated With ImmunoCult™ Human CD3/CD28 T Cell Activator
EasySep™-isolated human T cells were stimulated with ImmunoCult™ Human CD3/CD28 T Cell Activator and cultured in ImmunoCult™-XF T Cell Expansion Medium. Activation of viable CD3+ T cells was assessed by CD25 expression using flow cytometry. On day 0, the frequency of CD25 positive cells was (A) 5.6 ± 2.4% (mean ± SD). Following 3 days of culture, the frequency of CD25 positive cells was (B) 75.4 ± 13.8% (mean ± SD) when stimulated with ImmunoCult™ Human CD3/CD28 T Cell Activator.

Figure 3. Robust Human T Cell Expansion with ImmunoCult™ Human CD3/CD28 T Cell Activator
EasySep™-isolated human T cells were expanded over 12 days with ImmunoCult™ Human CD3/CD28 T Cell Activator in ImmunoCult™-XF T Cell Expansion Medium supplemented with Human Recombinant IL-2. On day 0, 1 x 10^6 EasySep™-isolated human T cells were stimulated with 25 μL of ImmunoCult™ Human CD3/CD28 T Cell Activator in ImmunoCult™-XF T Cell Expansion Medium supplemented with 10 ng/mL Human Recombinant IL-2. On days 3, 5, 7, and 10, viable cells were counted and fresh medium supplemented with IL-2 was added. No additional ImmunoCult™ Human CD3/CD28 T Cell Activator was added during the 12-day culture period (mean ± SD in 6 experiments with 3 donors).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (18)
Publications (5)
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Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.