產(chǎn)品名稱 |
K7M2 wt [K7M2-WT] |
商品貨號 |
B167172 |
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
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
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? |
Cellular Products |
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%.
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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.
- 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 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.
- 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 na?ve 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
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