| 產品名稱 |
L10BIOBR-MAPKK |
| 商品貨號 |
B198413 |
| Organism |
Mus musculus, mouse |
| Cell Type |
melanocyte |
| Product Format |
frozen |
| Morphology |
melanocyte |
| Culture Properties |
adherent |
| Biosafety Level |
2 Cells containing SV40 viral DNA sequences
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. |
| 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 Ref  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 |
| 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: - 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 gently pipetting.
-
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. - 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
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