ImmunoCult™-ACF Dendritic Cell Medium

Medium for the culture and differentiation of human monocytes into dendritic cells and their maturation

ImmunoCult™-ACF Dendritic Cell Medium

Medium for the culture and differentiation of human monocytes into dendritic cells and their maturation

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Medium for the culture and differentiation of human monocytes into dendritic cells and their maturation
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Overview

Reliably culture and differentiate human monocytes into dendritic cells (DCs) using the serum- and animal component-free (ACF) ImmunoCult™-ACF Dendritic Cell Medium.

For your convenience, the medium is available either individually or as part of ImmunoCult™ Dendritic Cell Culture Kit. Use the ImmunoCult™-ACF Dendritic Cell Medium in combination with ImmunoCult™-ACF Dendritic Cell Differentiation Supplement to generate DCs in vitro. To support the maturation of immature DCs to mature DCs, combine with the ImmunoCult™ Dendritic Cell Maturation Supplement.

For more information on protocols for culture and differentiation using ImmunoCult™ Dendritic Cell Medium, please explore the Product Information Sheet (PIS).
Subtype
Specialized Media, Supplements
Cell Type
Dendritic Cells, Monocytes
Species
Human
Application
Cell Culture, Differentiation
Brand
ImmunoCult
Area of Interest
Immunology
Formulation Category
Animal Component-Free, Serum-Free

Data Figures

Start: 54% CD4+CXCR3-CCR6+ T Cells

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.

Start: 54% CD4+CXCR3-CCR6+ T Cells

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).

Start: 54% CD4+CXCR3-CCR6+ T Cells

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.

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
Catalog #
10987, 10986
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
10987, 10986
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.