Description: The Immortalized Mouse CD4+ CD8+ T Cells (MOHITO) has characteristics of an immortalized cell line, but are not immortalized by any immortalization reagents. It expresses the Notch1 and Jak1 mutations, as well as TCR rearrangement, both of which are characteristics to human T-cell acute lymphoblastic leukemia (T-ALL). The cells are derived from CD4+ CD8+ T cells and are interleukin-7 (IL-7) dependent. The presence of IL-7 and IL-2 are required to activate the JAK/STAT signaling pathway. Transfection with BCR-ABL1 or mutant JAK1 transforms the cells to become IL-7 independent for proliferation. These cells are able to induce T-ALL-like disease when injected into healthy Balb/c mice. This cell line represents a novel model system to study T cell-specific protein signaling and inhibition mechanisms and oncogenic mutations, moreover they can be utilized in in vitro and in vivo pharmaceutical studies.
Depending on the culture conditions and number of passages, the cells can become more CD8 single positive and lose CD4 expression. If CD4/CD8 double positive cells are needed, users will need to sort the cells regularly to select for the desired immunophenotype.Immortalization Method: Spontaneous immortalizationBioSafety Level: IISpecies: MouseSource Organ: SpleenGrowth Properties: SuspensionMorphology: RoundPopulation Doubling: 25-35 hoursSeeding Density: 400,000 - 800,000 cells/ml; Recommended split ratio is 1:2 to 1:5Markers: N/ADonor Age: N/ADonor Gender: N/ADonor Ethnicity: N/APropagation: Grow cells in 6-well plate (P0100) with the following conditions. The base medium for this cell line is Prigrow II medium available at abm (TM002). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 20%, 10 ng/ml mouse IL-7 (Z200645), 5 ng/ml mouse IL-2 ( Z200137), and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
This cell line must be maintained at 400,000-800,000 cells/ml and should be split every 2-3 days. Cultures split lower than 300,000 cells/ml may result in slower cell propagation or a decrease in cell viability. When splitting the cells, avoid complete replacement of media. Instead retain approximately 1/5 of the conditioned media to maintain cell propagation speed and health.
It's highly recommended to retain minimum 20% conditioned medium in each passage to ensure growth and propagation of cells.
* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.Quality Control: 1) Flow cytometry to analyze CD4+ CD8+ phenotype; 2) PCR assay for T cell receptor expression; 3) Sequence analysis to identify point mutations in Notch1 and Jak1Shipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C