In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation:
Isolation date: 1998 (The K7M2 wt cell line was derived by intra-osseous injection of the K7 murine osteosarcoma cell line to the proximal tibia of a BALB/c mouse. The resultant primary tumor spontaneously metastasized to the lungs. A pulmonary metastasis was removed from the lung and then surgically implanted to a paraosteal tibial muscle flap of a naïve mouse. This cycle of implantation of pulmonary metastasis to paraosteal tibial muscle flap sites was repeated, resulting in the K7M2 primary tumor. This tumor was dissociated to a single cell suspension to create the K7M2 wt cell line)
Applications:
tumor model
Tumorigenic:
Yes
Comments:
Antigen Expression: CD31 + [PubMed:1315100]
Products: factor VIII [PubMed: 11315100]; integrin binding sialoprotein (BSP) [PubMed11315100]; biglyan [PubMed: 11315100]; decorrin [PubMed:1315100]; villin 2 (ezrin) [PubMed: 11325848].
Expression of bone sialoprotein, biglyan, decorrin, and osteopontin is suggestive of bone lineage cells.
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Subculturing:
Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
An inoculum of 6 X 10(3) to 8 X 10(3) viable cells/cm2 is recommended.
Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 5 X 10(3) and 3 X 10(5) cells/cm2 Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended Medium Renewal: Two to three times weekly
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Doubling Time:
31 hours
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
derivative:ATCC CRL-2837
References:
89453: Khanna C, et al. An orthotopic model of murine osteosarcoma with clonally related variants differing in pulmonary metastatic potential. Clin. Exp. Metastasis 18: 261-271, 2000. PubMed: 11315100
89454: Khanna C, et al. Metastasis-associated differences in gene expression in a murine model of osteosarcoma. Cancer Res. 61: 3750-3759, 2001. PubMed: 11325848
89468: Khanna C, et al. The membrane-cytoskeleton linker ezrin is necessary for osteosarcoma metastasis. Nat. Med. 10: 182-186, 2004. PubMed: 14704791
