ImmunoCult™ Human CD3/CD28 T Cell Activator

概述

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. 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 research applications. If you require reagents suitable for use in cell therapy manufacturing, ImmunoCult™ Human CD3/CD28 T Cell Activators (Catalog #100-0784) are produced under relevant GMPs.

Contains
• Anti-human CD3 monospecific antibody complex <br> • 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
Immunology, Cell Therapy Development

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).

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.

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

Workflow Stages for T Cell Research

Cell Sourcing

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Workflow Stages for T Cell Engineering

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Cell Cryopreservation

Workflow Stages for T Cell Therapy Development

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Resources and Publications

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.

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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.

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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.

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ImmunoCult™ Human CD3/CD28 T Cell Activator

产品号 #10971_C

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What Our Scientist Says

概述

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Protocols and Documentation

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Scientists Helping Scientists™

ImmunoCult™ Human CD3/CD28 T Cell Activator

产品号 #10971_C

产品优势

What Our Scientist Says

概述

Data Figures

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (22)

Publications (5)

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Abstract

Abstract

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