VMM917
CRL-3232 ™
- Product category
-
Human cells
- Cell type
- melanocyte
- Morphology
- Epithelial-like
- Disease
-
Melanoma; Stage IV
- Applications
-
3D cell cultureCancer researchToxicology
- 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
-
Drug screening
Development of targeted therapy
Development of combination therapy
Tumor vaccine development
Characteristics
- Growth properties
- Adherent
- Passage history
- Original deposit at third passage from primary culture.
- Derivation
- Derived from a subcutaneous recurrence. Established from digest of groin tumor metastasis in PETI Medium.
- Age
- 57 years
- Ethnicity
- White
- Gender
- Male
- Clinical data
- Primary site: Left shoulder (2005) (2008)
- Antigen expression
- GP100, Tyrosinase, MAGE-A1
- Comments
-
NRAS Mutation: Q61L Q61R
CDKN2A Mutation: R80 (One allele, (wt or unknown for the other))
BRAF wt
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 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%.
- Temperature
-
37°C
- Atmosphere
-
95% Air, 5% CO2
- Handling procedure
-
1 amp --> 1 T-75
Thaw ampoule in 37oC water bath for approximately 2 minutes. Transfer thawed cell suspension to a 15.0 mL centrifuge tube containing 9 mL complete medium. Mix suspension by gentle inversion. Remove 0.5-1.0 mL for cell count. Centrifuge remaining suspension in the 15 mL centrifuge tube at 175-195 x g (1000 rpm in an IEC HN SII centrifuge or equivalent) for 5 minutes, RT˚. Discard supernatant and gently resuspended pellet in 5 ml fresh complete medium. Transfer 5 ml re-suspended cells into 1 T-75 flask containing 10 ml fresh medium. Place the cells in a 5% CO2 incubator @ 37oC - Subculturing procedure
-
Volumes are for a T-75 flask; Adjust accordingly
- Remove and discard the cell culture medium from the flask.
- Rinse the cell monolayer with Dulbecco’s PBS without calcium or magnesium and remove.
- Add 3 to 4 ml of the trypsin-EDTA solution, rotate flask to rinse cell monolayer, remove trypsin solution, and incubate at 37oC.
- 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.
- Subculture as necessary.
- Place the flask back into the incubator. Examine the culture the following day to ensure the cells have reattached and are actively growing.
- Repeat when cells reach confluence.
- Culture maintenance
-
Cultures are grown @ 37°C in a 95% air, 5% CO2 environment.
Medium change every 2-4 days.
Cells produce heavy debris at higher confluence.
Cells are slow growing. - Reagents for cryopreservation
- Fetal bovine serum supplemented with 10% (v/v) DMSO (ATCC 4-X)
Quality control specifications
- Bacterial and fungal testing
- Not detected
- Population doubling capacity
- ≥ 15 in complete growth medium
- STR profiling
-
Amelogenin: X,Y
D5S818: 11
D13S317: 11
D7S820: 8,10
D16S539: 12
vWA: 18
TH01: 7,9.3
TPOX: 8,11
CSF1PO: 13
History
- Depositors
- Craig L. Slingluff, Jr. M.D.
- Year of origin
- 2008
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
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.
Images
Frequently Asked Questions
References
Curated Citations
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
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