L10BIOBR-GFP (ATCC® CRL-2770)

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 1
Age newborn
Strain B10.BR
Applications
Together, L10BIOBR-GFP (CRL-2770) and L10BIOBR-MAPKK (ATCC® CRL-2771) are valuable cell models for oncogenic transformation and signal transduction studies for melanoma [PubMed: 12514183].
The L10BIOBR-GFP cell line was derived as a negative control for CRL-2771. The immortalized murine melanocyte cell line L10BIOBR was transduced with pDIVA-GFP and subjected to puromycin selection.
Please note, although the L10BIOBR-GFP cell line harbors the gfp gene, as verified by PCR analysis, the cell line does not express sufficient GFP protein for detection of GFP fluorescence by flow cytometry or fluorescence microscopy.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
The L10BIOBR-GFP cell line was derived as a negative control for CRL-2771. The immortalized murine melanocyte cell line L10BIOBR was transduced with pDIVA-GFP and subjected to puromycin selection. Together, L10BIOBR-GFP (CRL-2770) and L10BIOBR-MAPKK (ATCC® CRL-2771) are valuable cell models for oncogenic transformation and signal transduction studies for melanoma [PubMed: 12514183]. Please note, although the L10BIOBR-GFP cell line harbors the gfp gene, as verified by PCR analysis, the cell line does not express sufficient GFP protein for detection of GFP fluorescence by flow cytometry or fluorescence microscopy.
Tumorigenic No
Effects
No, progressive tumor growth was not observed in nude mice
Comments
The L10BIOBR-GFP cell line was derived as a negative control for CRL-2771. The immortalized murine melanocyte cell line L10BIOBR was transduced with pDIVA-GFP and subjected to puromycin selection. Together, L10BIOBR-GFP (CRL-2770) and L10BIOBR-MAPKK (ATCC® CRL-2771) are valuable cell models for oncogenic transformation and signal transduction studies for melanoma [PubMed: 12514183]. Please note, although the L10BIOBR-GFP cell line harbors the gfp gene, as verified by PCR analysis, the cell line does not express sufficient GFP protein for detection of GFP fluorescence by flow cytometry or fluorescence microscopy.
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 2 X 10(4) cells/sq. cm.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 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 29 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

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Restrictions

This product's use is governed by the Limited Use License. For information on purchasing a license to use this product for research (for-profit entities) or commercial purposes (any entity) other than those permitted in the limited use label license, contact the Licensing Department, Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, California 92008. Phone (760) 603-7200 or outlicensing@lifetech.com

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