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 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.
Cell Type
Immortalized Cells
Biosafety Level
2
Depositor
VIB
Disclaimer
1. 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 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We 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. 5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Donor History
Balb/c Mouse
Expression Region
Human vWF and Human caveolin-1
Growth Properties
Suspension, round
Growth Conditions
Grow cells in 6-well plate (P0100) with the following conditions. PriGrow II (TM002) + 20% FBS + 1% Penicillin/Streptomycin Solution (G255) + 10ng/ml mouse IL-7 + 5ng/ml mouse IL-2 + 1ng/ml mouse IL-3, 37.0?, 5% CO?. It's highly recommended to retain minimum 20% conditioned medium in each passage to ensure growth and propagation of cells
Quantity
1x106 cells / 1.0 ml
Tissue
Blood
Morphology
Polygonal
Notes
1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use.To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism
Mouse (M. musculus)
Species
Mouse (M. musculus)
Propagation
Use of PriCoatTM T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 6%, Nu-Serum (Corning 355100) to a final concentration of 6% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Change media every 2-3 days.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.
Cryopreservation
Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control
RT-PCR was used to detect the presence of SV40 large T antigen and CD81
Applications Range
Research Use Only.
Shipping
Ship with dry ice.
Product Format
Frozen
Product Use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Storage
Vapor phase of liquid nitrogen, or below -130?.
Thawing Protocol
1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions.
Subculture Protocol
1. Simply add fresh complete media directly to the culture. Do not allow cell density to exceed 1x10? cells/ml. 2. Alternatively, replace complete growth media by centrifugation and re-suspend the cell pellet in fresh complete media, and add appropriate aliquots of the cell suspension to new culture vessels, as desired. 3. Incubate the cells at the recommended conditions.
Reviews of Immortalized Mouse CD4+ CD8+ T Cells (MOHITO)