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Print Version |
| BioSafety Level |
II |
| Organism |
CD11c:SV40LgT-transgenic C57BL/6 mice |
| Source Organ |
Spleen |
| Growth Properties |
Adherent |
| Morphology |
Small Aggregates |
| Recommended Seeding Density |
2.5 x 105 cells/cm2, should not be split lower than 5 x 104 cells per cm2 |
| Markers |
CD11c, CD24, MHC-II+, DEC205, Clec9A, B220, CD11b, IRF4, IRF8, CD4 |
| Applications |
For Research Use Only |
| Immortalization Method |
Isolated from C57BL/6 transgenic mice carrying SV40 Large T oncogene |
| Description |
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 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 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. T3034), TLR9 knockout (Cat. No. T3035) and Ifnar1 knockout (Cat. No. T3036) MutuDC, these dendritic cell lines provide a powerful tool in vaccine science and immunotherapy, particularly in strategizing target antigens to CD8α+ subset. |
| Procedure Overview |
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| Propagation |
We recommend to grow these cells on Poly-L-Lysine coated flask in which Poly-L-Lysine Coating Solution (0.1%) is available at abm (TM061).
The base medium for this cell line is PriGrow V, available at abm (TM015). To make the completed growth medium, add the following components to the base medium: 10% decomplemented fetal calf serum (PAN biotech; P40-37500), 50 µM β-mercaptoethanol, 1% HEPES and 1% Penicillin/Streptomycin (G255). 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.
IMPORTANT: This cell line is very sensitive to FBS batches. We strongly recommend end users to use FBS obtained from PAN biotech. |
| Preservation |
1. Freeze Medium: Complete growth medium with 50% decomplemented fetal calf serum (PAN biotech; P40-37500) and 10% DMSO.
2. Storage Temperature: Liquid nitrogen vapour phase. |
| Quality Control |
1) Direct antigen presentation and cross-antigen presentation were evaluated by the MHC-I (SIINFEKL/OT-I ) and MHC-II (OVA323-339/OT-II) restricted systems; 3) proteome profile and surface markers assessed by RT-PCR; RT-PCR; 3) IL-12 cytokine secretion analyzed using ELISA ; 4) Response to PAMP stimulation evaluated by functional assays. |
| Disclaimer |
1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.
2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).
3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.
4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. |
