A375-MA1
CRL-3222 ™
- Product category
-
Human cells
- Organism
-
Homo sapiens, human
- Morphology
- epithelial
- Tissue
- Skin
- Disease
-
Melanoma
- Applications
-
3D cell culture
- Product format
- Frozen
- Storage conditions
-
Vapor phase of liquid nitrogen
Documentation
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
Required Products
These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.
Detailed product information
General
- Specific 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.
Characteristics
- Volume
- 1.0 mL
- Growth properties
- Adherent
- 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.
- Age
- 54 years
- Gender
- Female
- Tumorigenic
- Yes
Handling information
- Unpacking and storage instructions
-
- Check all containers for leakage or breakage.
- Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until ready for use.
- Complete 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%.
- Temperature
-
37°C
- Atmosphere
-
95% Air, 5% CO2
- Subculturing procedure
-
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 - Reagents for cryopreservation
- Complete growth medium supplemented with 10% (v/v) DMSO (ATCC 4-X)
Quality control specifications
- STR profiling
-
Amelogenin: X
CSF1PO: 11,12
D13S317: 11,14
D16S539: 9
D5S818: 12
D7S820: 9
THO1: 8
TPOX: 8,10
vWA: 16,17
History
- Depositors
- R Hynes
- Year of origin
- 2004
Legal disclaimers
- Intended use
- This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
- Warranty
-
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
- Disclaimers
-
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.
While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.
This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.
Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
Permits & Restrictions
For-profit Research Use License from Massachusetts Institute of Technology
For-profit Organizations: For every order of this item, you must work directly with the contributor, Massachusetts Institute of Technology to (i) negotiate a research-use license, and/or (ii) have the contributor provide authorization to ATCC to ship this item under your existing license. We cannot ship this item until we receive communication directly from Massachusetts Institute of Technology that we are authorized to ship each order.
We are providing the following contact information, but this information may change without notice:
MIT Technology Licensing Office
One Cambridge Center, Kendall Square
Technology Licensing Office, NE18-501
Cambridge, Massachusetts 02142
Phone: 617-253-6966
Email: [email protected]
Once ATCC has received authorization from Massachusetts Institute of Technology, your order will be reviewed, and this item will be released for shipment if all requirements are met. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.
Import Permit for the State of Hawaii
If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.
Frequently Asked Questions
References
Curated Citations
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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