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ImmunoCult? Human CD3/CD28/CD2 T Cell Activator

Human T cell activation and expansion reagent

ImmunoCult? Human CD3/CD28/CD2 T Cell Activator

Human T cell activation and expansion reagent

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Human T cell activation and expansion reagent
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Product Advantages


  • Robust activation and expansion of human T cells without the use of magnetic beads, feeder cells, or antigen

  • Provides a gentle activation stimulus that maintains high viability of activated and expanded T cells

  • Highly stable, filter-sterilized soluble reagent

Overview

Achieve robust activation and expansion of T cells in the absence of magnetic beads, feeder cells, or antigens.

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. ImmunoCult? Human CD3/CD28/CD2 T Cell Activator consists of soluble antibody complexes that bind to and cross-link CD3, CD28, and CD2 cell surface ligands, thereby providing the required primary and co-stimulatory signals for T cell activation.

This product is designed for research applications. If you require reagents suitable for use in cell therapy manufacturing, ImmunoCult? Human CD3/CD28/CD2 T Cell Activators (Catalog #100-0785) are produced under relevant GMPs.
Contains
? Anti-human CD3 monospecific antibody complex
? Anti-human CD28 monospecific antibody complex
? Anti-human CD2 monospecific antibody complex
Subtype
Supplements
Cell Type
T Cells, T Cells, CD4+, T Cells, CD8+
Species
Human
Application
Activation, Cell Culture, Expansion
Brand
ImmunoCult
Area of Interest
Immunology, Cell Therapy Development

Data Figures

Activated Morphology of Human T Cells Stimulated With ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator

Figure 1. Activated Morphology of Human T Cells Stimulated With ImmunoCult™ Human CD3/CD28/CD2 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/CD2 T Cell Activator, and cultured in ImmunoCult™-XF T Cell Expansion Medium (Catalog #10981).

Activation of EasySep™ Isolated Human T Cells Stimulated With ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator

Figure 2. Activation of EasySep™ Isolated Human T Cells Stimulated With ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator

EasySep™-isolated human T cells were stimulated with ImmunoCult™ Human CD3/CD28/CD2 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) 88.8 ± 3.2% (mean ± SD) when stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator.

Robust Human T Cell Expansion with ImmunoCult? Human CD3/CD28/CD2 T Cell Activator

Figure 3. Robust Human T Cell Expansion with ImmunoCult? Human CD3/CD28/CD2 T Cell Activator

EasySep?-isolated human T cells were expanded over 12 days with ImmunoCult? Human CD3/CD28/CD2 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/CD2 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/CD2 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.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Name
Catalog #
10990, 10970
Lot #
All
Language
English
Document Type
Product Name
Catalog #
10990, 10970
Lot #
All
Language
English

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

Publications (29)

CD137L promotes immune surveillance in melanoma via HLTF regulation L. Liang et al. Nature Communications 2025 Sep

Abstract

Immune checkpoint blockers (ICBs) have demonstrated substantial efficacy across various malignancies, yet the benefits of ICBs are limited to a subset of patients. Therefore, it is essential to identify novel therapeutic targets. By integrating multi-omics data from cohorts of patients with melanoma treated with ICBs, a positive correlation is observed between tumor CD137L expression and the efficacy of PD-1 blockade. Functionally, CD137L induction in cancer cells significantly enhances anti-tumor immunity by promoting CD8 + T cell survival, both in vivo and in vitro. Mechanistically, helicase-like transcription factor (HLTF) is identified as a pivotal transcriptional regulator of CD137L , controlling its expression through phosphorylation of serine at position 398. Therapeutically, the AMPK agonist AICAR (acadesine) as an inducer of CD137L , exhibiting synergistic effects with PD-1 or CTLA-4 blockade. In summary, our findings elucidate a mechanism controlling CD137L expression and highlight a promising combination therapy to enhance the efficacy of ICBs in melanoma. One Sentence Summary: Inducing co-stimulatory immune checkpoint CD137L expression in melanoma cells enhances T cell-mediated anti-tumor immunity. Subject terms: Tumour immunology, Cancer immunotherapy
Comprehensive real-time metabolic profiling of peripheral blood mononuclear cells reveals important methodological considerations for immunometabolism research M. Macleod et al. Frontiers in Immunology 2025 Oct

Abstract

The utility of measuring real-time cellular bioenergetics of peripheral blood mononuclear cells (PBMCs) as biomarkers in disease monitoring, such as the bioenergetic health index, is of emerging interest. However, various experimental factors can impact the accuracy and reproducibility of these measurements. Methods: : PBMC bioenergetics were probed in real-time using extracellular flux analysis to identify optimal seeding density and injection protocol. Using a modified protocol, we assessed the extent to which blood processing time and isolation method (SepMate? vs. EasySep? Direct) influence PBMC bioenergetics under basal and stimulated conditions. Advanced metabolic control analysis including mitochondrial and glycolytic ATP supply flux, respiratory control ratio, bioenergetic health index, and mitochondrial toxicity index were used to identify and quantify PBMC bioenergetics. Results: Measures of metabolic profiling such as mitochondrial respiration, glycolytic activity, ATP supply flux, and respiratory control ratio were significantly diminished in PBMCs due to blood processing delay (48–72 hours) and were influenced by isolation method. Extended blood processing time significantly lowered T cell activation capacity in PBMCs, evidenced by decreased responses of mitochondrial and glycolytic ATP supply to CD3/CD28 activation. Discussion/Conclusion: This study demonstrates that key experimental variables including blood processing time and isolation method critically affect the reliability and biological relevance of PBMC metabolic assessments, highlighting the importance of protocol standardisation for accurate bioenergetic biomarker measurements.
Targeting triple-negative breast cancer using cord-blood CD34? HSPC-derived mesothelin-specific CAR-NKT cells with potent antitumor activity Li et al. Journal of Hematology & Oncology 2025 Oct

Abstract

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer characterized by the lack of ER, PR, and HER2 expression. Its aggressive behavior, high degree of tumor heterogeneity, and immunosuppressive tumor microenvironment (TME) are associated with poor clinical outcomes, rapid disease progression, and limited therapeutic options. Although chimeric antigen receptor (CAR)-engineered T cell therapy has shown certain promise, its applicability in TNBC is hindered by antigen escape, TME-mediated suppression, and the logistical constraints of autologous cell production. In this study, we employed hematopoietic stem and progenitor cell (HSPC) gene engineering and a feeder-free HSPC differentiation culture to generate allogeneic IL-15-enhanced, mesothelin-specific CAR-engineered invariant natural killer T ( Allo15 MCAR-NKT) cells. These cells demonstrated robust and multifaceted antitumor activity against TNBC, mediated by CAR- and NK receptor-dependent cytotoxicity, as well as selective targeting of CD1d + TME immunosuppressive cells through their TCR. In both orthotopic and metastatic TNBC xenograft models, Allo15 MCAR-NKT cells demonstrated potent antitumor activity, associated with robust effector and cytotoxic phenotypes, low exhaustion, and a favorable safety profile without inducing graft-versus-host disease. Together, these results support Allo15 MCAR-NKT cells as a next-generation, off-the-shelf immunotherapy with strong therapeutic potential for TNBC, particularly in the context of metastasis, immune evasion, and treatment resistance. The online version contains supplementary material available at 10.1186/s13045-025-01736-9.