L10BIOBR-MAPKK細胞
ATCC Number： CRL-2771?
數(shù)量： 大量
品系： B10.BR
運輸方式： 凍存運輸
細胞形態(tài)： 其他
細胞類型： 其他細胞類型
年限： newborn
是否是腫瘤細胞： 0
物種來源： 小鼠
生長狀態(tài)： 貼壁生長
Designations: L10BIOBR-MAPKK
Depositors: JL Arbiser
L10BIOBR-MAPKK細胞Biosafety Level: 2 [Cells containing SV40 viral DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Mus musculus
Morphology: melanocyte
Source: Cell Type: melanocyte;
Strain: B10.BR
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: Isolation date: January 1, 2002
Applications: tumor model
Tumorigenic: Yes
Age: newborn
Comments: L10BIOBR-MAPKK細胞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].
Propagation: ATCC complete growth medium: Ham's F10 medium supplemented with 50 ng/ml TPA (Sigma Catalogue No. P-8139) and 7% horse serum
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
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: L10BIOBR-MAPKK細胞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
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 24 hours
Related Products: recommended serum:ATCC 30-2040
derived from same cell line:ATCC CRL-2770
References: 89472: Govindarajan B, et al. L10BIOBR-MAPKK細胞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
