The temporomandibular joint (TMJ) is the jaw joint that hinges to allows complex movements related to eating and swallowing. TMJ disc tissue permits mandibular movement and its degeneration has been linked to many clinical pathologies related to joint function, including crepitus, arthritis and headdaches. TMJ disc tissue comprises of a complex matrix of cells with regional difference in morphology, including chondrocyte -like cells and fibroblast-like cells. This organization of cells allow for the disspation of forces acting on the jaw joint. Immortalized Mouse TMJ Disc Fibroblast-Like Cells were derived from fibroblast-like cells from a 12-week-old female mouse. These cells are designed to be suitable for cell-type specific studies on the physiology and progression of TMJ disc disorders. Characterization was conducted by Western blot, qRT-PCR, and immunofluorescence.
Cell Type
Immortalized Cells
Biosafety Level
2
Depositor
University of Michigan
Disclaimer
1. All test parameters provided in the CoA are conducted using abm's standardized culture system and 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 order placement. 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. 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 or any other non-RUO application(s). 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are 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".
Use of PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow VIII (TM018) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?
Quantity
1x106 cells / 1.0 ml
Tissue
Bone
Morphology
Small aggregates
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 IMDM (1x) + GlutamaxTM (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 beta-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 0.22µm before use.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.To heat-inactivate 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. Heat-inactivated FBS can be stored at -20°C for long term. Avoid frequent freeze-thaw cycling.
Cryopreservation
Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control
1) Flow cytometry; 2) ELISA; 3) qPCR
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
Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions.
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