| 產(chǎn)品名稱 |
A375-MA1 |
| 商品貨號(hào) |
B229912 |
| Organism |
Homo sapiens, human |
| Tissue |
skin |
| Product Format |
frozen 1.0 mL |
| Morphology |
epithelial |
| 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 |
malignant melanoma |
| Age |
54 years |
| Gender |
female |
| Applications |
This cell line is useful to study the mechanisms of metastasis. It has been used with microarray analyses to identify metastasis-specific genes using a functional genomics approach and in proteomics analyses. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
This increased metastatic cell line was derived using an in vivo selection process of highly metastatic cells from a population of poorly metastatic tumor cells, A375 (ATCC CRL-1619). The A375-M1 (ATCC CRL-3222) cell line was derived by i.v. injection of A375 cells into nude mice. Lung metastases were harvested and amplified in vitro as cell lines. The A375-M1 cell line was reinjected into mice for a second round of selection, lung metastases harvested and amplified in vitro as A375-M2 (ATCC CRL-3223) cells. The A375-M1 and A375-M2 cell lines were transfected with a plasmid containing the ecotropic receptor for murine retrovirus and selected for neomycin resistance. This is useful for RNAse protection assays. |
| Clinical Data |
54 years
female |
| Tumorigenic |
yes |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
|
| Subculturing |
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (ATCC 30-2200) or 0.05% (w/v) Trypsin-0.02% EDTA solution (ATCC PCS-999-003) 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.
6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Volume |
1.0 mL |
| STR Profile |
Amelogenin: X
CSF1PO: 11,12
D13S317: 11,14
D16S539: 9
D5S818: 12
D7S820: 9
THO1: 8
TPOX: 8,10
vWA: 16,17 |
| Name of Depositor |
R Hynes |
| Year of Origin |
2004 |
| Year of Deposit |
2014 |
| References |
Clark EA, et al. Genomic analysis of metastasis reveals an essential role for RhoC. Nature 406: 532-535, 2000. PubMed: 10952316
Lei X, et al. Gene Expression Changes in an Animal Melanoma Model Correlate with Aggressiveness of Human Melanoma Metastases. Mol. Cancer Res. 6(5): 760-769, 2008. PubMed: 18505921
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