Immortalized Mouse Dendritic Cells (MutuDC1940)永生化小鼠樹突狀細胞

Cat No.: NTCC925761

As messengers bridging the innate and adaptive immune system, the conventional tissue-resident DC (cDC) acts as sentinels in secondary lymphoid organs and other tissues for antigen capture and presentation. The Wild-type NTCC? MutuDC1940 cell line is an immortalized splenic CD8α+ subset (CD11chigh,B220-,DEC205+,CD24high,CD11b-) of conventional dendritic cells derived from the CD11c:SV40LgT transgenic mice.

The NTCC? MutuDC1940 cell line retains response to TLR ligands such as CpG (TLR9-L) and PolyIC (TLR3-L) and to a lesser extent LPS (TLR4-L) stimulation by up-regulation of co-stimulatory molecules CD40, CD80 and CD86. In addition to responding to PAMP stimulation and producing Th1 cytokines such as IL-12, these cells are also capable of presenting antigen in the context of both MHC-I and MHC-II, including direct antigen presentation and cross-presentation of cell-associated antigens. Together with TLR3 knockout (Cat. No. NTCC-T3034), TLR9 knockout (Cat. No. NTCC-T3035) and Ifnar1 knockout (Cat. No. NTCC-T3036) MutuDC, these dendritic cell lines provide a powerful tool in vaccine science and immunotherapy, particularly in strategizing target antigens to CD8α+ subset.

Image:

Immortalization Method: Isolated from C57BL/6 transgenic mice carrying SV40 Large T oncogene

BioSafety Level: II

Species: Mouse

Source Organ: Spleen

Growth Properties: Adherent

Morphology: Small aggregates

Population Doubling: 45 - 55 hours

Seeding Density: 20,000 – 60,000 cells/cm2.

Recommended split ratio:no greater than 1:3.

Markers: CD11c, CD24, MHC-II+, DEC205, Clec9A, B220, CD11b, IRF4, IRF8, CD4

Propagation: The base medium for this cell line is IMDM (1x) + GlutamaxTM (NTCC-31980-030). To make the complete growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum (TM999) to a final concentration of 10%, 1% of 7.5% Sodium Bicarbonate Solution, 50 μM β-mercaptoethanol, HEPES to a final concentration of 10 mM, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Filter the complete media at 0.22μm before use.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.To decomplement FBS:1. Let the FBS bottle completely thaw overnight at 4°C. 2. Leave the FBS bottle in water bath for 30 minutes at 56°C. 3. Decomplemented FBS can be stored at -20°C for long term. Avoid frequent freeze-thaw cycling.Change the complete media every 2-3 days. Do not let media colour change to yellow.

To thaw NTCC? MutuDC1940:1. Thaw the vial in 37oC water bath until there is no more than a small cube of ice.2. Transfer the cells to a 15ml tube containing 5ml of pre-warmed complete media. 3. Centrifuge the cells at 290xg for 5 minutes. 4. Carefully discard supernatant without disturbing the cell pellet and gently resuspend the cells in 1ml of complete media by lightly pipetting up and down. 5. Seed the cells at 20,000 - 60,000 cells/cm2.To subculture NTCC? MutuDC1940: 1. It is recommended to subculture when cells are at 70-90% confluency. 2. Aspirate old media. Some cells in suspension are still viable cells. Alternatively, the supernatant containing the suspended cells can also be collected and centrifuged (Skip to Step 6 in protocol). 3. Add 1:1 ratio of 1X sterile PBS and 0.25% Trypsin-EDTA (NTCC-TM051). 4. Incubate cells at room temperature for 3-5 minutes and agitate the culture vessel until 90% ofthe cells have detached. 5. Immediately neutralize the trypsin by adding complete media equal to the volume of trypsin + PBS added. 6. Collect the cells and centrifuge at 290xg for 5 minutes. 7. Discard the supernatant and gently resuspend the cells in complete media by lightly pipetting up and down. 8. Recommended split ratio is no more than 1:3. Stimulation can be performed using PolyIC (5 μg/ml), CpG (2 mM) or LPS (5 μg/ml). These cells are especially sensitive to FBS requirement, thus, it is advised to use the same batch for culturing cells that show best result in supporting their culture. We also recommend the addition of extra 1% Hepes to the complete media to encourage propagation of the cells.

To freeze NTCC? MutuDC1940: Recommended freezing medium: Complete growth medium with decomplemented FBS to a final concentration of 50% and 5% DMSO. Storage temperature: Liquid nitrogen vapour phase.

Name：Immortalized Mouse Dendritic Cells (MutuDC1940)
中文名稱：永生化小鼠樹突狀細胞(MutuDC1940)
Organism種屬：
Tissue組織來源:
Cell Type細胞類型:
Morphology形態:
Culture Properties培養特性:
Disease疾病:
Biosafety Level生物安全級別:
Age年齡:
Gender性別:
Applications應用:
Images圖片:
Complete Growth Medium完全培養基:
Subculturing傳代方法:
Cryopreservation凍存方法:Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions培養條件:Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
STR Profile鑒定數據:
References參考文獻:

Supplier供應商:BioVector質粒載體菌株細胞蛋白抗體基因保藏中心
NTCC典型培養物保藏中心
Tel電話：+86-010-53513060，400-800-2947
Email：Biovector@163.com
http://www.biovector.net