L10BIOBR-MAPKK (ATCC® CRL-2771)

Organism: Mus musculus, mouse  /  Cell Type: melanocyte  / 

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Organism Mus musculus, mouse
Cell Type melanocyte
Product Format frozen
Morphology melanocyte
Culture Properties adherent
Biosafety Level 2 Cells containing SV40 viral DNA sequences
Age newborn
Strain B10.BR
Applications
tumor model
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
The L10BIOBR-MAPKK cell line (ATCC CRL-2771) was derived by infecting the immortalized murine melanocyte cell line, L10BIOBR, with pBABE which encodes a constitutively active MAPKK. The vector contains the SV40 viral DNA sequences and the puromycin resistance gene. The cells were selected in medium containing puromycin. The introduction of the MAPKK gene into melanocytes leads to tumorigenesis in nude mice, activation of the angiogenic switch and increased production of the proangiogenic factor, vascular endothelial growth factor (VEGF), and matrix metalloproteinases (MMPs). Activation of MAP kinase signaling may be an important pathway involved in melanoma transformation. Inhibition of MAP kinase signaling may be useful in the prevention and treatment of melanoma. The L10BIOBR-MAPKK cell line and the corresponding negative control, L10BIOBR-GFP (CRL-2770), are a model for melanoma tumorigenesis and signal transduction [PubMed: 12514183].
Tumorigenic Yes
Effects
Yes, produces tumors in nude mice inoculated subcutaneously with 10(6) cells
Comments
The L10BIOBR-MAPKK cell line (ATCC CRL-2771) was derived by infecting the immortalized murine melanocyte cell line, L10BIOBR, with pBABE which encodes a constitutively active MAPKK. The vector contains the SV40 viral DNA sequences and the puromycin resistance gene. The cells were selected in medium containing puromycin. The introduction of the MAPKK gene into melanocytes leads to tumorigenesis in nude mice, activation of the angiogenic switch and increased production of the proangiogenic factor, vascular endothelial growth factor (VEGF), and matrix metalloproteinases (MMPs). Activation of MAP kinase signaling may be an important pathway involved in melanoma transformation. Inhibition of MAP kinase signaling may be useful in the prevention and treatment of melanoma. The L10BIOBR-MAPKK cell line and the corresponding negative control, L10BIOBR-GFP (CRL-2770), are a model for melanoma tumorigenesis and signal transduction [PubMed: 12514183].
Complete Growth Medium Ham's F10 medium supplemented with 50 ng/ml TPA (Sigma Catalogue No. P-8139) and 7% horse 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 5 X 10(3) to 7 X 10(3) viable cells/sq. cm. is recommended.
  6. Incubate cultures at 37°C.
Interval: Subculture when cells reach a concentration of 4 X 10(4) cells/sq. cm.
Subcultivation Ratio: A subcultivation of 1:6 to 1:8 is recommended
Medium Renewal: Two to three times weekly
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: 37.0°C
Population Doubling Time 24 hours
Name of Depositor JL Arbiser
Year of Origin January 1, 2002
References

Govindarajan B, et al. Malignant transformation of melanocytes to melanoma by constitutive activation of mitogen-activated protein kinase kinase (MAPKK) signaling. J. Biol. Chem. 278: 9790-9795, 2003. PubMed: 12514183

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

Govindarajan B, et al. Malignant transformation of melanocytes to melanoma by constitutive activation of mitogen-activated protein kinase kinase (MAPKK) signaling. J. Biol. Chem. 278: 9790-9795, 2003. PubMed: 12514183