ImmunoCult™

Activation, Expansion and Maintenance of Immune Cells

Advance your research with ImmunoCult™ products designed to ensure optimal activation, expansion and differentiation of the immune cells of your interest. These specialized media, activators and supplements allow you to culture immune cells under defined stimulatory conditions for consistent and reliable results.

ImmunoCult™ for T Cells

Advance your basic T cell immunology and clinical T cell engineering research with ImmunoCult™ products designed for:

  • Robust T cell activation and expansion without the use of magnetic beads, feeder cells or antigens.
  • Consistent T cell expansion in serum- and xeno-free expansion medium.

ImmunoCult™ for Dendritic Cells

Streamline your research on dendritic cells (DCs) with ImmunoCult™ products designed for:

  • Generation of mature DCs from isolated monocytes.
  • Obtaining a high yield of mature DCs with the desired phenotype and function.

Why Use ImmunoCult™?

  • Serum-free and animal-component-free medium.
  • Ready-to-use and optimized formulation for activation, expansion or differentiation of immune cells.
  • Results in a high yield of cells with the desired phenotype and function without the need to supplement with serum.
  • Media, activators and supplements can be used on their own or combined.

ImmunoCult™ for T Cells

ImmunoCult™
Human T Cell Activators


Application:

Activation and Expansion of T Cells

Description:

Soluble antibody complexes that recognize and cross-link CD3 and co-stimulatory molecules on the surface of the T cell in the absence of magnetic beads or feeder cells. It can be used in combination with ImmunoCult™-XF T Cell Expansion Medium or any other media for culturing human T cells.

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

ImmunoCult™
Human T Cell Expansion Medium


Application:

Expansion of T Cells

Description:

Serum-free and xeno-free medium optimized for the in vitro culture and expansion of human T cells isolated from peripheral blood. It can be used in combination with ImmunoCult™ T Cell Activators for bead-free activation of T cells.

Advantages:

  • No need to supplement the medium with serum
  • Supports robust T cell expansion at levels comparable to those in serum-containing media
  • Maintains similar proportions of CD4+/CD8+ cells to the start of culture after expansion
  • Expanded T cells are able to produce cytokines including IFN-gamma and IL-4
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New Partnership Enabling Development and Manufacturing of T Cell Immunotherapies

GE Healthcare has partnered with STEMCELL Technologies Inc. to provide T-Cell reagents for commercial-scale cell therapy production. Under the agreement, GE Healthcare will commercialize cGMP-grade versions of STEMCELL Technologies’ T-Cell reagents for the isolation, activation, and culture of T-cells in clinical applications.

Read the Press Release

Data

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

Figure 1. T cells are activated when stimulated with ImmunoCult™ Human CD3/CD28 or CD3/CD28/CD2 T Cell Activator.

EasySep™ isolated 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 2. T cells exhibit an activated morphology when activated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator.

Images of EasySep™-isolated 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 3. T cells show robust expansion when cultured in ImmunoCult™-XF T Cell Expansion Medium.

T cells were expanded over 14 days with ImmunoCult™ Human CD3/CD28 T Cell Activator or ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator 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 4. T cells remain viable after 14 days of culture in ImmunoCult™-XF T Cell Expansion Medium.

T cells were expanded over 14 days with ImmunoCult™ Human CD3/CD28 T Cell Activator or ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator 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.)

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

Figure 5. T cells expanded in ImmunoCult™-XF T Cell Expansion Medium have similar CD4/CD8 composition as T cells at the start of culture.

T cells were expanded over 21 days with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator in ImmunoCult™-XF T Cell Expansion Medium supplemented with rhIL-2. At day 0 and day 21, T cells were harvested and analyzed for (A) CD4+ and (B) CD8+ expression. (Note that T cells were reactivated during the course of expansion every 7 days.)

Figure 6. T cells expanded in ImmunoCult™-XF T Cell Expansion Medium are able to functionally produce intracellular IFN-gamma and IL-4.

T cells were expanded over 21 days with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator in ImmunoCult™-XF T Cell Expansion Medium supplemented with rhIL-2. At day 21, T cells were harvested and analyzed for intracellular IFN-gamma and IL-4 after stimulation with PMA and ionomycin for 4 hours. (Note that cells were reactivated during the course of expansion every 7 days.)



ImmunoCult™ for Generation of Dendritic Cells

ImmunoCult™ Dendritic Cell Medium

Application:

Culture, differentiation and maturation of dendritic cells (DCs)

Description:

Serum-free and animal-component free (ACF) medium optimized for the in vitro culture and differentiation of human monocytes into dendritic cells. It can be used in combination with ImmunoCult™ DC Differentiation and Maturation Supplements.

Advantages:

  • No need to supplement the medium with serum
  • Supports DC differentiation and maturation
  • Can be used on its own or combined with differentiation and maturation supplements

ImmunoCult™ Differentiation Supplement

Application:

Differentiation of monocytes to immature DCs

Description:

Contains a combination of animal component-free (ACF) recombinant human cytokines formulated to support the differentiation of immature DCs from human monocytes.

Advantages:

  • Defined formulation that has been standardized and optimized
  • Promotes consistent differentiation of monocytes into immature dendritic cells
  • The differentiation supplement is ready-to-use and is simply added to the medium

ImmunoCult™ Maturation Supplement

Application:

Maturation of immature to mature DCs

Description:

Formulated to support the maturation of immature dendritic cells.

Advantages:

  • Induces maturation of immature dendritic cells
  • Ready-to-use and optimized formulation
  • Obtain high yields of mature DCs
  • Mature DCs produce cytokines and induce T cell proliferation
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Data

Figure 1. Protocol Diagram.

Mature DCs were generated by culturing EasySep™ isolated monocytes at 1 x 106 cells/mL in ImmunoCult™-ACF Dendritic Cell Medium (Catalog #10987) with added ImmunoCult™-ACF Dendritic Cell Differentiation Supplement (Catalog #10988). At day 3, the medium with differentiation supplement was replaced and cells were incubated for 2 more days. At day 5, without changing the medium, ImmunoCult™ Dendritic Cell Maturation Supplement (Catalog #10989) was added to the culture. At day 7, fully mature DCs were harvested for downstream applications.

Figure 2. Mature DCs generated with ImmunoCult™-ACF Dendritic Cell Medium with Supplements show desired phenotype.

EasySep™ isolated monocytes were cultured and differentiated into mature DCs as described in Figure 1. (A) The percentage of CD14 and CD83 expression in cells at day 7 (mature DCs) was determined by flow cytometry. At day 7, a total of 93 ± 5% of the cells expressed the mature DC marker CD83 and only 1 ± 1% of cells still expressed the monocyte marker CD14 (mean ± SD, n=39). Yield of mature DCs was determined by count of total viable cells at day 7 relative to the count of viable monocytes used for initial culture at day 0. At day 7, the yield of viable mature DCs corresponded to 45 ± 25% (mean ± SD, n=39). (B) Immature DCs were cultured as described in Figure 1. At day 5, cells were cultured with maturation supplement for 2 days (mature DCs) or without maturation supplement (immature DCs). Supernatant was collected at day 7 and IL-12p70 levels were determined by ELISA. Concentrations of IL-12p70 in supernatant of mature and immature DCs were 361 ± 81 and 5 ± 2 pg/mL, respectively (mean ± SEM, n=27).

Figure 3. Mature DCs generated with ImmunoCult™-ACF Dendritic Cell Medium and Supplements induce T cell proliferation.

Mature DCs generated with ImmunoCult™-ACF Dendritic Cell Medium and Supplements (ImmunoCult) or other serum-free competitor media (competitor 1 and 2) and corresponding supplements when applicable (competitor 2), were cultured in ImmunoCult™-XF T Cell Expansion Medium with 1 x 105 CFSE labeled (A) allogeneic CD3+ T cells (MLR assay) or (B) autologous CD8+ T cells (antigen-specific T cell response). (A) Cells were cultured at a DC:T cell ratio of 1:25. (B) Prior to culture with T cells, immature DCs were loaded with HLA Class I peptides derived from the human Cytomegalovirus, Epstein-Barr Virus and Influenza Virus (CEF peptide pool) and stimulated with maturation supplement for 2 days. Cells were cultured at a DC:T cell ratio of 1:4 or 1:10. (A,B) CFSE labeled T cells were incubated in media alone (negative control) or with ImmunoCult™ Human CD3/CD28 T Cell Activator (positive control). After 5-7 days in culture the number of dividing T cells ( CD3+CFSElo) was assessed by flow cytometry (mean ± SEM) (A) n=5 (B) n=4 (competitor 1 and 2, n=3). Mature DCs generated in ImmunoCult™-ACF Dendritic Cell Medium induced proliferation of allogeneic and antigen-specific T cells similar to DCs generated in either competitor media. Competitors 1 and 2, include in no particular order, CellGro DC Medium (CellGenix) and PromoCell DC Generation Medium DXF.

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