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

Our collaboration with GE Healthcare aims to give researchers the confidence of a path to the clinic with cGMP-grade T cell reagents. 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.

Products

The following human T cell isolation, activation and expansion reagents aim to be developed as cGMP-grade reagents, and we can currently work with you to qualify these reagents under an approved investigational new drug (IND) or clinical trial application (CTA).

EasySep™ Release CD3 Positive Selection Kit

The EasySep™ Release Human CD3 Positive Selection Kit is designed to positively-select particle-free CD3+ cells from fresh or previously frozen PBMCs or washed leukapheresis samples using a column-free immunomagnetic selection. Obtain up to 99% pure CD3+ cells free of magnetic particles in under 30 minutes, that are immediately ready for downstream applications, such as T cell activation and expansion.

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.

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.

Try these isolation, activation and expansion reagents in your lab. Request your free sample.

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).


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