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VMM5A
VMM5A
規(guī)格:
貨期:
編號(hào):B167207
品牌:Mingzhoubio

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產(chǎn)品名稱 VMM5A
商品貨號(hào) B167207
Organism Homo sapiens, human
Tissue Melanoma, Lymph Node Metastasis
Cell Type Melanocyte
Product Format frozen
Morphology Epithelial-like
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 Melanoma, Stage IIIC; malignant
Age 81
Gender Male
Ethnicity Caucasian
Applications Drug screening
Development of targeted therapy
Development of combination therapy
Tumor vaccine development
Storage Conditions liquid nitrogen vapor phase
Images ATCC CRL-3226 Cell Micrograph
Derivation Derived from tumors taken from tumor-involved lymph nodes from patients at the University of Virginia
Clinical Data Primary Site: Right Posterior Shoulder; Metastatic Site: Lymph Node, Right Neck
Antigen Expression Positives: High VEGF-R2, Low GP100, Tyrosinase, Low MAGE-A1, MMP-1
HLA Typing A2.1,A1101,B39,B44, BW4,BW6,C7(17)DR7, DR8,DR11,DQ2,DQ7
Comments NRAS: wt
CDKN2A: wt
BRAF Mutation: V600E
APC Mutation
PDGFRA Mutation: D842V
PTEN Mutation: R173H 
more sensitive to BAY43-9006 (BRAF kinase inhibitor) and to
rapamycin (mTOR kinase inhibitor), compared to cell lines with wild-type B-Raf

Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
Subculturing

Volumes are for a T-75 flask; Adjust accordingly

  1. Remove and discard the cell culture medium from the flask.
  2. Rinse the cell monolayer with Dulbecco’s PBS without calcium or magnesium and remove.
  3. Add 3 to 4 ml of the trypsin-EDTA solution, rotate flask to rinse cell monolayer, remove trypsin solution, and incubate at 37oC.
  4. Once the cells appear to be detached, add 10 ml of complete growth medium with a pipette to the cell suspension to inactivate the trypsin. Gently wash any remaining cells from the growth surface of the   flask. Check the  cells with the microscope to be sure that most (>95%) are single cells. If cell clusters are apparent, continue to disperse the cells with gentle pipetting.
  5. Subculture as necessary.
  6. Place the flask back into the incubator. Examine the culture the following day to ensure the cells have reattached and are actively growing.
  7. Repeat when cells reach confluence.
Cryopreservation Fetal bovine serum, 90%; DMSO, 10%
Culture Conditions Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2 ), 5%
STR Profile Amelogenin: X,Y
D5S818: 9
D7S820: 9,10
D13S317: 12
D16S539: 12
vWA: 16
TH01: 9.3
TPOX: 8,11
CSF1PO: 11
Sterility Tests

Pass

Name of Depositor Craig L. Slingluff, Jr. M.D.
Passage History Unknown. 2 passages from 2003 frozen cell line stock, but it is not known how long the line was in culture after being established from tumor tissue obtained in 1992.
Year of Origin 1992
References

Slingluff C, et al. Melanomas with concordant loss of multiple melanocytic differentiation proteins: immune escape that may be overcome by targeting unique or undefined antigens. Cancer Immunol. Immunother. 48:661-672, 2000. PubMed: 10752474

Molhoek K, et al. Human melanoma cytolysis by combined inhibition of mammalian target of rapamycin and Vascular Endothelial Growth Factor/Vascular Endothelial Growth Factor Receptor-2. Cancer Res. 68: (11), 2008. PubMed: 18519701

Molhoek K, et al. Comprehensive analysis of RTK activation in human melanomas reveals autocrine signaling through IGF-1R. Melanoma Res 21(4): 274–284, 2011. PubMed: 21654344

Molhoek K, et al. Synergistic inhibition of human melanoma proliferation by combination treatment with B-Raf inhibitor BAY43-9006 and mTOR inhibitor Rapamycin. J. Trans. Med. 3(39): 2005. PubMed:16255777

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