K7M2 wt [K7M2-WT] (ATCC® CRL-2836)

Organism: Mus musculus, mouse  /  Cell Type: osteoblast  /  Tissue: bone; derived from metastatic site: lung  /  Disease: osteosarcoma

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Organism Mus musculus, mouse
Tissue bone; derived from metastatic site: lung
Cell Type osteoblast
Product Format frozen
Morphology osteoblast
Culture Properties adherent
Biosafety Level 1
Disease osteosarcoma
Strain BALB/c
Applications
This cell line is a suitable tumor model.
Storage Conditions liquid nitrogen vapor phase
Antigen Expression

CD31 + RefKhanna 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 

Genes Expressed secreted phosphoprotein 1 (osteopontin) 

factor VIII RefKhanna 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 

integrin binding sialoprotein (BSP) RefKhanna 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 

biglyan RefKhanna 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 decorrin RefKhanna 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

villin 2 (ezrin) RefKhanna C, et al. Metastasis-associated differences in gene expression in a murine model of osteosarcoma. Cancer Res. 61: 3750-3759, 2001. PubMed: 11325848 

Tumorigenic Yes
Effects
Yes, forms tumors in BALB/c mice with spontaneous metastasis to the lungs in over 90% of mice inoculated
Comments Expression of bone sialoprotein, biglyan, decorrin, and osteopontin is suggestive of bone lineage cells.
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%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by pipetting gently. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 6 x 103 to 8 x 103 viable cells/cm2 is recommended.
  5. Incubate culture vessels at 37°C. We recommend that you maintain cultures at a cell concentration between 5 x 104 and 3 x 105 cells/cm2.

Subcultivation Ratio: 1:4 to 1:6
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells: a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Population Doubling Time 31 hours
Name of Depositor C Khanna
Year of Origin 1998
References

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

Khanna C, et al. Metastasis-associated differences in gene expression in a murine model of osteosarcoma. Cancer Res. 61: 3750-3759, 2001. PubMed: 11325848

Khanna C, et al. The membrane-cytoskeleton linker ezrin is necessary for osteosarcoma metastasis. Nat. Med. 10: 182-186, 2004. PubMed: 14704791

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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

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

Khanna C, et al. Metastasis-associated differences in gene expression in a murine model of osteosarcoma. Cancer Res. 61: 3750-3759, 2001. PubMed: 11325848

Khanna C, et al. The membrane-cytoskeleton linker ezrin is necessary for osteosarcoma metastasis. Nat. Med. 10: 182-186, 2004. PubMed: 14704791

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