K7M2-pCI Neo (ATCC® CRL-2837)

Organism: Mus musculus, mouse  /  Cell Type: osteoblast  /  Disease: osteosarcoma

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Organism Mus musculus, mouse
Cell Type osteoblast
Product Format frozen
Morphology osteoblast
Culture Properties adherent
Biosafety Level 2 Cells containing CMV and SV40 viral DNA sequences
Disease osteosarcoma
Strain BALB/c
Applications
tumor model
Images
Derivation
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. 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 [PubMed:11315100]. Expression of bone sialoprotein, biglyan, decorrin, and osteopontin is suggestive of bone lineage cells.
Genes Expressed
decorin,secreted phosphoprotein 1 (osteopontin),villin 2 (ezrin),Expression of bone sialoprotein, biglyan, decorrin, and osteopontin is suggestive of bone lineage cells.
Cellular Products
decorin
secreted phosphoprotein 1 (osteopontin)
villin 2 (ezrin)
Tumorigenic Yes
Effects
Yes, forms tumors in BALB/c mice with spontaneous metastasis to the lungs in over 90% of mice inoculated
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.
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 [PubMed:11315100]. Expression of bone sialoprotein, biglyan, decorrin, and osteopontin is suggestive of bone lineage cells.
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 0.8 mg/ml G-418 and 10% fetal bovine serum
Subculturing
Protocol:
  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 gently pipetting.
  5. 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/sq. cm is recommended.
  6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 3 X 10(4) and 1 X 10(5) cells/sq. cm
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Two to three times weekly
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Population Doubling Time 12 hours
Name of Depositor C Khanna
Year of Origin 2004
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-pCI Neo cell line was derived from the K7M2 wt (ATCC CRL-2836) cell line. K7M2 wt cells were transfected with the pCI Neo vector. The vector carries the human cytomegalovirus (CMV) immediate-early enhancer/promoter region to promote constitutive expression of cloned DNA inserts in mammalian cells. It also contains SV40 viral sequences and the neomycin resistance gene.

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
Other 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-pCI Neo cell line was derived from the K7M2 wt (ATCC CRL-2836) cell line. K7M2 wt cells were transfected with the pCI Neo vector. The vector carries the human cytomegalovirus (CMV) immediate-early enhancer/promoter region to promote constitutive expression of cloned DNA inserts in mammalian cells. It also contains SV40 viral sequences and the neomycin resistance gene.