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ImmunoCult™ Human CD3/CD28 T Cell Activator

Human T cell activation and expansion reagent

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ImmunoCult™ Human CD3/CD28 T Cell Activator

Human T cell activation and expansion reagent

From: 179 USD
Catalog #
10971_C
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

What Our Scientist Says

We want to make it easier to activate and expand human T cells while still maintaining a high viability of these cells. That's why we developed ImmunoCult™ Human CD3/CD28 T Cell Activator.

Jessie YuScientist
Jessie Yu, Scientist

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. 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.
Contains
• Anti-human CD3 monospecific antibody complex
• Anti-human CD28 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
Cell Therapy Research, Immunology

Data Figures

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

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).

Activation of Human T Cells stimulated With ImmunoCult™ Human CD3/CD28 T Cell Activator

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.

Robust Human T Cell Expansion 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.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
ImmunoCult™ Human CD3/CD28 T Cell Activator
Catalog #
10971, 10991
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
ImmunoCult™ Human CD3/CD28 T Cell Activator
Catalog #
10971, 10991
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.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (18)

T Cell Reagents for Your Cellular Therapy Research
Brochure
T Cell Reagents for Your Cellular Therapy Research
Isolate Human Immune Cells
Brochure
Isolate Human Immune Cells
Optimization of Human T Cell Expansion Protocol: Effects of Early Cell Dilution
Technical Bulletin
Optimization of Human T Cell Expansion Protocol: Effects of Early Cell Dilution
Production of Chimeric Antigen Receptor T Cells
Wallchart
Production of Chimeric Antigen Receptor T Cells
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
The Immune Response to HIV Poster
Wallchart
The Immune Response to HIV Poster
CAR T Cell Manufacturing Workflow: Isolation, Activation and Expansion
0:59
Video
CAR T Cell Manufacturing Workflow: Isolation, Activation and Expansion
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
1:37
Video
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep...
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
4:30
Video
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
1:57
Video
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Qualification of Ancillary/Raw Materials for Clinical Use
54:39
Webinar
Qualification of Ancillary/Raw Materials for Clinical Use
T Cell Differentiation and Cancer Immunity
48:32
Webinar
T Cell Differentiation and Cancer Immunity
Online Immunology Journal Club: Expanding the Therapeutic Tool Kit for CAR T Cells
47:14
Webinar
Online Immunology Journal Club: Expanding the Therapeutic Tool Kit for CAR T Cells
Workflow Solutions for Human T Cell Isolation and Expansion
Scientific Poster
Workflow Solutions for Human T Cell Isolation and Expansion
Rapid Expansion of Functional Human T Cells Using a Novel Serum-Free and Xeno-Free Culture Medium
Scientific Poster
Rapid Expansion of Functional Human T Cells Using a Novel Serum-Free and Xeno-Free Culture Medium
Optimization of Human T Cell Activation and Expansion Protocols Improves Efficiency of Genetic Modification and Overall Cell Yield
Scientific Poster
Optimization of Human T Cell Activation and Expansion Protocols Improves Efficiency of Genetic Modif...

Publications (5)

Competition between PAF1 and MLL1/COMPASS confers the opposing function of LEDGF/p75 in HIV latency and proviral reactivation. R. Gao et al. Science advances 2020 may

Abstract

Transcriptional status determines the HIV replicative state in infected patients. However, the transcriptional mechanisms for proviral replication control remain unclear. In this study, we show that, apart from its function in HIV integration, LEDGF/p75 differentially regulates HIV transcription in latency and proviral reactivation. During latency, LEDGF/p75 suppresses proviral transcription via promoter-proximal pausing of RNA polymerase II (Pol II) by recruiting PAF1 complex to the provirus. Following latency reversal, MLL1 complex competitively displaces PAF1 from the provirus through casein kinase II (CKII)-dependent association with LEDGF/p75. Depleting or pharmacologically inhibiting CKII prevents PAF1 dissociation and abrogates the recruitment of both MLL1 and Super Elongation Complex (SEC) to the provirus, thereby impairing transcriptional reactivation for latency reversal. These findings, therefore, provide a mechanistic understanding of how LEDGF/p75 coordinates its distinct regulatory functions at different stages of the post-integrated HIV life cycles. Targeting these mechanisms may have a therapeutic potential to eradicate HIV infection.
View publication
Gut-Liver Physiomimetics Reveal Paradoxical Modulation of IBD-Related Inflammation by Short-Chain Fatty Acids. M. Trapecar et al. Cell systems 2020 mar

Abstract

Although the association between the microbiome and IBD and liver diseases is known, the cause and effect remain elusive. By connecting human microphysiological systems of the gut, liver, and circulating Treg and Th17 cells, we created a multi-organ model of ulcerative colitis (UC) ex vivo. The approach shows microbiome-derived short-chain fatty acids (SCFAs) to either improve or worsen UC severity, depending on the involvement of effector CD4 T cells. Using multiomics, we found SCFAs increased production of ketone bodies, glycolysis, and lipogenesis, while markedly reducing innate immune activation of the UC gut. However, during acute T cell-mediated inflammation, SCFAs exacerbated CD4+ T cell-effector function, partially through metabolic reprograming, leading to gut barrier disruption and hepatic injury. These paradoxical findings underscore the emerging utility of human physiomimetic technology in combination with systems immunology to study causality and the fundamental entanglement of immunity, metabolism, and tissue homeostasis.
View publication
A Simple and Scalable Strategy for Analysis of Endogenous Protein Dynamics. M. K. Schwinn et al. Scientific reports 2020 jun

Abstract

The ability to analyze protein function in a native context is central to understanding cellular physiology. This study explores whether tagging endogenous proteins with a reporter is a scalable strategy for generating cell models that accurately quantitate protein dynamics. Specifically, it investigates whether CRISPR-mediated integration of the HiBiT luminescent peptide tag can easily be accomplished on a large-scale and whether integrated reporter faithfully represents target biology. For this purpose, a large set of proteins representing diverse structures and functions, some of which are known or potential drug targets, were targeted for tagging with HiBiT in multiple cell lines. Successful insertion was detected for 86{\%} of the targets, as determined by luminescence-based plate assays, blotting, and imaging. In order to determine whether endogenously tagged proteins yield more representative models, cells expressing HiBiT protein fusions either from endogenous loci or plasmids were directly compared in functional assays. In the tested cases, only the edited lines were capable of accurately reproducing the anticipated biology. This study provides evidence that cell lines expressing HiBiT fusions from endogenous loci can be rapidly generated for many different proteins and that these cellular models provide insight into protein function that may be unobtainable using overexpression-based approaches.
View publication
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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.

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