MPC 862L
CRL-3540 ™
MPC 862L is a cell line isolated from a mouse pheochromocytoma.
- Organism
-
Mus musculus, mouse
- Classification
- Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
- Morphology
- Epithelial like and/or rounded
- Tissue
- Adrenal gland
- Disease
-
Pheochromocytoma
- Applications
-
Cancer research
- 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
-
Pheochromocytoma research
Neuroendocrine research
NF1 LOH Research
High-throughput screening
Phenylethanolamine-N-methyltransferase ·
Transcription Studies
RET signaling Studies
Characteristics
- Cells per vial
- 6.0 x 106
- Comments
-
CRL-3540 is a pheochromocytoma developed from heterozygous Nf1 knockout mice. This cell line was established from irradiated 10 day old pups of a F1 male from 129SV (Nf1+/Nf1n31) male crossed with wild-type C57BL/6 females. CRL-3538 offers an alternative model to PC12 cells (CRL-1721). CRL-3538 differs from CRL-1721 as they show little or no expression of the NGF receptor TrkA and do not respond to NGF. (Powers et al., 2000).
The cells will produce phenyl-L-methly transferase (PNMT)messenger RNA and low levels of protein when 10 uM dexamethasone is added to culture per the cell line depositor. The cells express a high amount of Ret tyrosine kinase receptor.
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, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium:
- heat-inactivated horse serum to a final concentration of 10%
- fetal bovine serum to a final concentration of 5%
- Temperature
-
37°C (35-37°C)
- Atmosphere
-
95% Air, 5% CO2
- Subculturing procedure
-
Before initiating subculture flasks must be coated with 50 µg/ mL collagen I from rat tail (Thermofisher Scientific catalog #: A1048301 or equivalent). It is recommended to follow the vendor instructions for thin coating procedure.
- It is recommended to subculture at or before 80% confluence. Cells may not reach 100% confluence and grow in clumps which will become necrotic if not passaged.
- Before beginning subculture procedure prepare a solution of 0.25% Gibco™ Trypsin (ThermoFisher Scientific catalog # 15050- 065) + 80 μg/mL Deoxyribonuclease I. Deoxyribonuclease I (DNase I) should be added to an aliquot of 0.25% Gibco™ Trypsin fresh prior to use.
- To prepare 10 mg/mL DNase I stock solution, aseptically combine: o 100 mg Roche™ DNase I (Sigma catalog # 11284932001) o 10 mL Molecular Grade Water (ATCC catalog # 60-2450) Add 8 μL stock DNase I per 1 mL 0.25% Trypsin immediately prior to use.
- Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with PBS to remove all traces of serum that contains trypsin inhibitor.
-
Add 2.0 to 3.0 mL of 0.25% Gibco™ Trypsin (ThermoFisher Scientific catalog # 15050-065) + 80 μg/mL Deoxyribonuclease I) to flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 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. - Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Centrifuge the cell suspension at approximately 150 to 400 x g for 8 to 12 minutes to remove dissociation agent.
- Resuspend the cell pellet in an appropriate amount of complete culture medium
- If cell clumps are present then clumps should be disrupted using a 5 mL serological pipette followed by a P1000.
-
Add appropriate aliquots of the cell suspension to new culture vessels.
Cultures can be established between 1.0 x 1054 and 2.06.0 x 105 viable cells/cm2. - Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 1.0 X 1054 and 2.0 X 105 cell/cm2.Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommendedMedium Renewal: Every 2 to 3 days - Reagents for cryopreservation
-
Gibco™ Recovery™ Cell Culture Freezing Media (ThermoFisher Scientific catalog # 12648-010).
Quality control specifications
- Bacterial and fungal testing
- Not detected
- Mycoplasma contamination
- Not detected
- Virus testing
-
Cytomegalovirus (CMV): Not detectedEpstein-Barr virus (EBV): Not detectedHepatitis B virus (HBV): Not detectedHuman Immunodeficiency virus (HIV): Not detectedHuman papillomavirus (HPV): Not detected
History
- Deposited as
- Mus musculus
- Depositors
- Dr. Arthur S. Tischler
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.
Frequently Asked Questions
Powered by Bioz
For product-related inquiries and issues, contact Product Experience:
Hours of Operation
Monday - Friday
9:00am - 5:00pm
US Eastern Time
US Eastern Time
