ImmunoCult™树突状细胞成熟添加物

用于人单核细胞来源树突状细胞成熟的添加物

产品号 #(选择产品)

产品号 #10989_C

用于人单核细胞来源树突状细胞成熟的添加物

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总览

使用ImmunoCult™树突状细胞成熟添加物,将未成熟树突状细胞(DC)转化为成熟DC。

该产品作为ImmunoCult™树突状细胞培养试剂盒的组分,也可单独购买,以方便您的使用。ImmunoCult™树突状细胞成熟添加物专为搭配ImmunoCult™-ACF树突状细胞培养基和ImmunoCult™-ACF树突状细胞分化添加物使用而设计,用以支持未成熟DC的成熟过程。

关于使用ImmunoCult™树突状细胞成熟添加物进行DC成熟的详细方法,请参阅产品说明书(PIS)。

亚型
专用培养基,添加剂
 
细胞类型
树突状细胞(DCs),单核细胞
 
种属

 
应用
细胞培养,分化
 
品牌
ImmunoCult
 
研究领域
免疫
 
制剂类别
Animal Component-Free,无血清
 

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 #
10989
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
10989
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.

更多信息

更多信息
Species Human
Formulation Category Animal Component-Free, Serum-Free
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