CellGenix® TCM GMP-Prototype

T Cell Medium

CellGenix® TCM is an optimized, xeno-free medium used for serum-free expansion and differentiation of (CD3+) T cells. It promotes an early onset of T cell expansion and is thereby optimal for short protocols and a faster cell therapy manufacturing process. CellGenix® TCM enables both CD4+ and CD8+ T cell expansion. It supports a central memory and early differentiated memory T cell phenotype. Additionally, it boosts a high proportion of cytokine producing cells including polyfunctional cells.

General Information

CellGenix® TCM is optimized for serum-free expansion and cultivation of T cells. It has been optimized and verified with CAR T cell protocols. In addition, it supports expansion of other cell types such as Treg and iNKT cells.

We offer the cytokines and growth-factors needed for optimal growth separately. This gives you the freedom to customize and create your own optimal cytokine mixture.

GMP-Prototype grade

We initially offer our CellGenix® TCM as GMP-Prototype. This allows you to test the T Cell Medium before our GMP grade will be available. We are currently generating supplementary quality control data, including extended real-time stability data, in order to meet our GMP quality standards. The performance is identical to the future GMP TCM, allowing for a seamless transition to GMP.

name
CellGenix® T Cell Medium GMP-Prototype
Quality grade
GMP-Prototype
Cell Type
T cells
CellGenix® TCM GMP-Prototype
volume
format
Order Number
500 ml
bottle
24814 -0500

Would you like to test if CellGenix® TCM is the right T cell medium for your T cell therapy product?

Contact your local sales manager


Infographic
–  How to choose the best T cell medium for your CAR-T cell manufacturing process

CellGenix® TCM GMP-Prototype

Features

  • Serum-free and xeno-free medium
  • Early onset of T cell expansion and sustained viability
  • Supports expansion of CD4+ and CD8+ T cells
  • Promotes a central memory and early differentiated memory T cell phenotype
  • High proportion of cytokine producing cells including polyfunctional cells

Application

  • Serum-free expansion and cultivation of T cells
  • Optimized for and verified with CAR T cells
  • Supports expansion of other cell types such as Treg and iNKT cells

Technical Details

Osmolality
280-320 (mOsm/kg H2O), determined according to Ph. Eur.
pH
6.9 – 7.3, determined according to Ph. Eur.
Endotoxin
≤ 0.5 EU/ml, determined according to Ph. Eur.
Sterility
Sterility test of final product determined according to Ph. Eur.
Mycoplasma
Mycoplasma PCR-test according to Ph Eur.
Functionality assay
Ex vivo-culture of primary human CD3+ T-Cells
Intended use
For preclinical ex vivo use. Not intended for human in vivo application.

Handling Instructions

Shipment

Ambient temperature

Storage & Stability

Store at +2°C to +8°C. Please protect from light.

 

Primary Packaging Details

Medium Bottle

  • Material of bottle: Copolymer PET (Polyethylene Terephthalate), Food Grade Certification
  • Material of bottle top: High-Density Polyethylene (HDPE), Food Grade Certification

Documents

Static culture in 96-well U-bottom

Protocol: CD3+ T cells were purified from healthy human donors (n=6; EasySep™ Human T Cell Isolation Kit, Stemcell Technologies)*. Cells were activated with Dynabead Human T-Activator CD3/CD28 (Thermo Fischer) at a ratio of 1:1 bead:cell in CellGenix® TCM or competitor media in the presence of IL-7 and IL-15 (10 ng/ml, CellGenix). Media and IL-7 and IL-15 (10 ng/ml, CellGenix) were added on d2, d4, d7 and d9. Cells were splitted on d4, d7 and d9.

*In some experiments, cells were frozen in CellGenix® TCM with 10% DMS after purification and thawed before activation.

High T cell expansion with CellGenix TCM

Figure 1: Exponential fit of T cell expansion kinetics
CD3+ T cells purified from human donors (n=6) were activated using anti-CD3/anti-CD28-coupled
magnetic beads in CellGenix TCM or commercially available media for T cell expansion in the
presence of IL-7 and IL-15 (10 ng/ml, CellGenix).

   

Early onset of T cell expansion and sustained viability throughout serum-free culture

Figure 2: Fold T cell expansion and cell viability on the indicated day of culture
CD3+ T cells were cultivated as described. On the indicated day cells were labeled with 7-AAD and viability was determined by flow cytometry. Median and spread of n=6 donors are displayed for expansion (upper panels) and cell viability (lower panels).

   

Robust expansion of CD4+ and CD8 + T cells

Figure 3: Proportions of CD4+ and CD8+ T cells
CD3+ T cells were cultivated as described. On the indicated day expression of CD4+ and CD8+ was determined by flow cytometry, n=6.

 
   

Culture supports an early memory phenotype (TN / TSCM)

 

Figure 4: Memory phenotype of starting population and after expansion in CellGenix TCM
CD3+ T cells were cultivated as described. On day 0 (left panel) and day 11 (right panel) the expression of memory cell markers was determined by flow cytometry for three donors. TCM, central memory T cell; TN, naïve T cell; TSCM, memory stem T cell; TEM, effector memory T
cells; TEMRA, terminally differentiated effector T cells.

   

High percentage of cytokine producing and particularly polyfunctional cells

Figure 5: Cytokine expression after expansion in CellGenix TCM
CD3+ T cells were cultivated as described. On day 11 of culture cells were restimulated using PMA and Ionomycin in the presence of BrefeldinA. Cells were fixed and permeabilized and expression of the indicated cytokines was determined by flow cytometry. Median and spread of n=6 donors are displayed for CD4+ (upper panels) and CD8+ (lower panels), black dots indicate statistical outliers.


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