ImmunoCult™ Dendritic Cell Culture Kit

Complete kit for differentiation of human monocytes into dendritic cells

ImmunoCult™ Dendritic Cell Culture Kit

Complete kit for differentiation of human monocytes into dendritic cells

From: 735 USD
Catalog #
(Select a product)
Complete kit for differentiation of human monocytes into dendritic cells
Add to Wish List

Product Advantages


  • Animal component-free and serum-free medium

  • No need to supplement the medium with serum

  • Optimized for differentiation of CD14+ monocytes into immature and subsequently mature DCs

  • High yields of mature DCs with the desired phenotype

  • Mature DCs are functional, produce the pro-inflammatory cytokine IL-12, and induce T cell proliferation

What's Included

• ImmunoCult™-ACF Dendritic Cell Medium, 100 mL (Catalog #10987)
• ImmunoCult™-ACF Dendritic Cell Differentiation Supplement, 1 mL (Catalog #10988)
• ImmunoCult™ Dendritic Cell Maturation Supplement, 0.5 mL (Catalog #10989)

Overview

Culture and differentiate human monocytes into dendritic cells (DCs) using the animal component-free (ACF) ImmunoCult™ Dendritic Cell Culture Kit.

For your convenience, the kit includes all the necessary components for you to generate active mature DCs from human monocytes in 7 days:

ImmunoCult™-ACF Dendritic Cell Medium for the in vitro culture and differentiation of human monocytes into DCs.
ImmunoCult™-ACF Dendritic Cell Differentiation Supplement to support the differentiation of immature DCs from human monocytes.
ImmunoCult™ Dendritic Cell Maturation Supplement to support the maturation of immature human DCs.

For more information on protocols for culture and differentiation using ImmunoCult™ Dendritic Cell Culture Kit, please explore the Product Information Sheet (PIS).
Contains
ImmunoCult™-ACF Dendritic Cell Medium contains only recombinant proteins and synthetic components
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 #
10985
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
10985
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
10985
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
10985
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.