Immortalized Mouse CD4+ CD8+ T Cells (MOHITO) | T0131

abm | Immortalized Mouse CD4+ CD8+ T Cells (MOHITO) | T0131

+ 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 immortalization

BioSafety Level:

II

Organism:

Balb/c Mouse

Species:

Mouse

Source Organ:

Spleen

Organ Type:

Blood

Growth Properties:

Suspension

Morphology:

Round

Passage Number:

N/A

Population Doubling:

25-35 hours

Seeding Density:

400,000 - 800,000 cells/ml; Recommended split ratio is 1:2 to 1:5

Markers:

N/A

Donor Age:

N/A

Donor Gender:

N/A

Donor Ethnicity:

N/A

Propagation:

Grow cells in 6-well plate with the following conditions. The base medium for this cell line is Prigrow II medium available at abm . To make the completed growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 20%, 10 ng/ml mouse IL-7 (Z200645), 5 ng/ml mouse IL-2 ( Z200137), and Penicillin/Streptomycin Solution to a final concentration of 1%.
Carbon dioxide (CO₂): 5%, Temperature: 37.0°C.

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 Jak1

Shipping Condition:

Dry Ice

Storage Condition:

liquid nitrogen or -180C