Immortalized Mouse Tumour Dendritic Cells (CD4⁻ MutuDC2) was isolated from the spleen of Batf3-/- CD11c-SV40LgT transgenic mice in C57BL/6JOlaHsd background. The CD4⁻ MutuDC2s display high levels of CD11c and MHC-II with lack of the pDC-specific markers B220, Gr-1 and PDCA-1. In their resting state they are CD80
hiand CD86
hi. These observations demonstrate that the CD4⁻ MutuDC2s have a relatively mature splenic resident conventional DC1 phenotype. They can be easily manipulated with standard experimental techniques and display great stability of their phenotypic and functional properties in long term culture as long as their passage number is maintained below 40-50. For these reasons the CD4⁻ MutuDC2s represent a new valuable tool for the study of dendritic cell biology.
Immortalization Method: Spontaneous SV40LgT-mediated tumorigenic transformation of splenic conventional Dendritic Cells in CD11c-SV40LgT-transgenic mice.
BioSafety Level: II
Organism: Mouse
Species: Mouse
Source Organs: Spleen
Organ Type: Spleen
Growth Properties: Adherent
Morphology: Round
Seeding Density: 72-144 hours
Markers: CD11c+, MHC-II+, F4/80lo, B220-, GR-1-, PDCA-1-, CD80+, CD86+, CD11b+, CD172a+, CD4-, CD8a-, CLEC9A-, FLT3+
Propagation: The base medium for this cell line is IMDM (1x) + Glutamax
TM (Gibco Ref: 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 0.22 µm before use.Carbon dioxide (CO
2): 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.
Quality Control: 1) Flow cytometry; 2) ELISA; 3) qPCR
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C
Reference: Pigni, M., Ashok, D., Stevanin, M., & Acha-Orbea, H. (2018). Establishment and Characterization of a Functionally Competent Type 2 Conventional Dendritic Cell Line. Frontiers in immunology, 9, 1912. https://doi.org/10.3389/fimmu.2018.01912