T Cell Therapy

From Bench to Bedside

Ready to take your T cell therapy research from bench to bedside? STEMCELL Technologies has entered a collaboration with GE Healthcare aiming to further enable development and manufacturing of T cell therapy with cGMP-grade T cell isolation, activation and expansion reagents.

Production of CAR T Cells Wallchart

Free Nature Protocols Wallchart summarizing the processes involved in producing CAR T cells for therapy.

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Immunology Feature: CAR T Cells

A mini-review of our selection of key advancements in CAR T cell therapy research.

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T Cell Isolation, Activation and Expansion

Optimize your human T cell research and manufacturing with the following isolation, activation and expansion reagents.

EasySep™ Release CD3 Positive Selection Kit

Isolate up to 99% pure CD3+ cells from fresh or previously frozen PBMCs or washed leukapheresis samples using a column-free immunomagnetic selection. Obtain viable and functional CD3+ cells that are free of magnetic particles and immediately ready for downstream applications in under 30 minutes.

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ImmunoCult™ Human CD3/CD28 T Cell Activator
ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator

Soluble antibody complexes that recognize and cross-link CD3 and co-stimulatory molecules for robust activation and expansion of human T cells without the use of magnetic beads, feeder cells or antigen. These activators provide a gentle activation stimulus that maintains high viability and are not exclusively licensed for use in T cell immunotherapy manufacturing.

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ImmunoCult™-XF T Cell Expansion Medium

Serum-free and xeno-free medium optimized for the rapid and consistent expansion of human T cells. This medium, with no serum supplementation needed, supports robust T cell expansion at levels comparable to serum-containing media and eliminates performance variability.

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See the data below and try these isolation, activation and expansion reagents in your lab.



For Cell Therapy Research Applications

Our collaboration with GE Healthcare aims to develop these T cell isolation, activation and expansion products into cGMP-grade reagents to give researchers the confidence of a path to the clinic. These products are designed for cell therapy research applications following the recommendations of USP <1043> on Ancillary Materials.

Why Use STEMCELL’s Reagents for Cell Therapy Research Applications?

  • Traceability documentation including CoAs and CoOs to help reduce time in preparing IND submissions or clinical trial applications.
  • Defined formulations to minimize lot-to-lot variability.
  • Extensive QC testing.
  • Experienced global professionals to help navigate regulatory processes.

We can currently work with you to qualify these reagents under an approved investigational new drug (IND) or clinical trial application (CTA). Read this technical bulletin to learn more about ancillary material qualification.


Contact us for more information on how we can support your cellular therapy research.

Integrated workflow for the manufacturing of chimeric antigen receptor (CAR) T cells using STEMCELL's reagents that aim to be cGMP-grade.

Isolation of T cells using (A) EasySep™ Release CD3 Positive Selection Kit; T cell activation using soluble (B) ImmunoCult™ Human T Cell Activator and gene modification to express the CAR construct; and expansion of CAR T cells in xeno-free and serum-free (C) ImmunoCult™-XF T Cell Expansion Medium.

Data

T cells isolated using EasySep™ Release Human CD3 Positive Selection Kit are highly purified.

Figure 1. Typical EasySep™ Release Human CD3 Positive Selection.

Starting with a single-cell suspension of human PBMCs, the CD3+ cell content of the isolated fraction is typically 98.7 ± 0.9% (mean ± SD using the purple EasySep™ Magnet). In the above example, the purities of the start and final isolated fractions are 38.4% and 99.0%, respectively.

T cells express activated phenotype and morphology when stimulated with ImmunoCult™ T Cell Activators in ImmunoCult™-XF T Cell Expansion Medium.

Figure 2. T cells are activated when stimulated with ImmunoCult™ Human CD3/CD28 or CD3/CD28/CD2 T Cell Activator.

T cells were cultured on day 0 with either ImmunoCult™ Human CD3/CD28 T Cell Activator (Catalog #10971) or ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator (Catalog #10970) in ImmunoCult™-XF T Cell Expansion Medium (Catalog #10981). Cells were gated on CD4+ T cells and CD8+ T cells and T cell activation was accessed by CD25+ expression on day 0 and day 3. At the start of culture, the CD25+ cell population was (A) 5.63 ± 2.4% (mean ± SD). After three days of activation, the CD25+ cell population was (B) 75.4 ± 13.8% (mean ± SD) when activated with ImmunoCult™ Human CD3/CD28 T Cell Activator and (C) 88.8 ± 3.2% (mean ± SD) when activated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator.

Figure 3. T cells exhibit an activated morphology when activated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator.

Images of T cells activated with (A) soluble ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator and (B) the competitor's bead-based activation reagent.

T cells show robust fold expansion and high viability when expanded with ImmunoCult™ T Cell Activators in ImmunoCult™-XF T Cell Expansion Medium.

Figure 4. T cells show robust expansion when stimulated with ImmunoCult™ Human T Cell Activators in ImmunoCult™-XF T Cell Expansion Medium.

T cells were expanded over 14 days with ImmunoCult™ Human CD3/CD28 T Cell Activator, ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator or Competitor's bead-based activation reagent in ImmunoCult™-XF T Cell Expansion Medium supplemented with rhIL-2. Fold expansion was determined between 0 to 14 days. (Note that T cells were not reactivated during the course of expansion.)

Figure 5. T cells stimulated with ImmunoCult™ Human T Cell Activators are highly viable.

T cells were expanded over 14 days with ImmunoCult™ Human CD3/CD28 T Cell Activator, ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator or Competitor's bead-based activation reagent in ImmunoCult™-XF T Cell Expansion Medium supplemented with rhIL-2. % viability was determined between 0 to 14 days. (Note that T cells were not reactivated during the course of expansion.)

Figure 6. ImmunoCult™-XF T Cell Expansion Medium supports greater T cell expansion than other serum-free and serum-supplemented media.

T cells were activated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator, and cultured in (A) ImmunoCult™-XF T Cell Expansion Medium or serum-free competitor media with rhIL-2 in three replicate cultures per donor, or cultured in (B) ImmunoCult™-XF T Cell Expansion Medium or serum-supplemented competitor media with rhIL-2 in three replicate cultures per donor. T cells were stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator on day 0 and every 7 to 8 days for the duration of the culture. T cells were analyzed on day 21 for fold expansion relative to the initial cell seeding density.
(A) Compared to all serum-free competitor media tested, ImmunoCult™-XF T Cell Expansion Medium showed significantly higher expansion of total T cells. Competitors 1 to 6 represent serum-free competitor media. Each column with error bars represents the mean ± S.E.M. (p<5x10-13 for ImmunoCult™-XF T Cell Expansion Medium versus all other serum-free media, tested using the linear mixed effect model with linear regression, n = 4 to 19 donors).
(B) compared to all serum-supplemented competitor media tested, ImmunoCult™-XF T Cell Expansion Medium showed similar or significantly higher expansion of total T cells. Competitors 1 to 4 represent serum-supplemented competitor media. Each column with error bars represents the mean ± s.e.m. (p<0.0006 for ImmunoCult™-XF T Cell Expansion Medium versus all other serum-supplemented media except for competitor 4, tested using the linear mixed effect model with linear regression, n = 1 to 19 donors).


Try these isolation, activation and expansion reagents in your own lab.



Complete Your T Cell Workflow

Human T Cell Isolation Kits

Isolate specific human T cell subsets or pan T cells for your particular T cell therapy research. Choose from a wide range of positive and negative selection cell isolation kits.

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CryoStor® CS10 Cryopreservation Medium

Cryopreserve human T cells in serum-free, animal component-free and defined freezing medium containing 10% dimethyl sulfoxide (DMSO). cGMP-manufactured with USP-grade components, CryoStor® CS10 provides a protective environment for cells during the freezing, storage and thawing processes.

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