CAR47
CRL-3235 ™
- Product category
-
Human cells
- Organism
-
Homo sapiens, human
- Cell type
- fibroblast
- Morphology
- fibroblast-like
- Tissue
- Adrenal gland
- Disease
-
Primary Pigmented Adrenocortical Disease PPNAD And Carney Complex
- 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.
Cells contain SV40 sequences
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 an immortalized cell line bearing a naturally occurring PRKA1A-inactivating mutation that is associated with multiple tumor formation derived from the adrenal tissue of a patient with Carney complex. PRKAR1A is a tumor suppressor gene. Applications include PKA associated characterization of cAMP-mediated mechanisms of cell cycle, signaling, proliferation and endocrine tumorigenesis.
Characteristics
- Growth properties
- Adherent
- Derivation
- The CAR47 cell line was derived from the adrenal gland that was removed from a patient diagnosed with PPNAD and Carney complex. After the first three to four passages, all cells in the growing cell lines had a fibroblastoid appearance, whereas both polygonal adrenocortical cells and fibroblasts were apparent at the beginning. At passage 10, the cells were transduced with a recombinant lentivirus expressing the large T antigen of SV40. The cell line was further cultured for several passages in the presence of blasticidin to ensure maintenance of the integrated lentivral cassette. In addition to the large T antigen, the integrated cassette also contains the gene that confers blasticidin resistance. The cells were initially frozen at passage 20.
- Age
- adult
- Gender
- Female
- Comments
-
The CAR47 cell line was derived from an adrenal gland that was removed from a patient diagnosed with PPNAD and Carney complex. This is an immortalized cell line with a naturally-occurring inactivating mutation in PRKAR1A, the regulatory subunit type 1A (R1alpha) of protein kinase A (PKA), which is associated with tumor formation.PKA isozyme balance is critical for the control of cAMP signaling and related cell cycle and proliferation changes. Aberrant cAMP signaling has been linked to adrenocortical and other, mostly endocrine, tumors. Inactivating mutations in the PRKAR1A gene are a known cause of Carney complex - an autosomal dominant multiple neoplasia syndrome associated with skin, heart, and other myxomas and a variety of endocrine tumors.
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 DMEM Medium Catalog No. 30-2002. To make the
complete growth medium, add the following components to the base medium:
- 10.0 µg/mL Blasticidin
- Temperature
-
37°C
- Atmosphere
-
95% Air, 5% CO2
- Subculturing procedure
-
Volumes used in this protocol are for 75 cm2 flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.05% (w/v) Trypsin - 0.02% EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- Add 1.0 to 2.0 mL of 0.05% 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 37.0°C to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
- Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
Medium Renewal: 2 to 3 times per week
Seeding Density: 1.0 x 104 to 3.0 x 104 cells/cm2 - Reagents for cryopreservation
- Complete growth medium supplemented with 20% (v/v) fetal bovine serum and 10% DMSO (ATCC 4-X)
Quality control specifications
- Functional tests
- Validation of PRKA1A-inactivating mutation
- STR profiling
-
Amelogenin: X
CSF1PO: 10
D13S317: 12,14
D16S539: 11
D5S818: 11,12
D7S820: 8,11
TH01: 7,9.3
TPOX: 8,11
vWA: 17,18
History
- Depositors
- M. Nesterova
- Year of origin
- 2005
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
Nesterova M, et al. An immortalized human cell line bearing a PRKAR1A-inactivating mutation: Effects of overexpression of the wild-type allele and other protein kinase A subunits. J. Clin. Endocrinol. Metab. 93: 565-571, 2008. PubMed: 18056771
Kirschner LS, et al. Genetic heterogeneity and spectrum of mutations of the PRKAR1A gene in patients with the Carney complex. Human Mol. Gen. 9: 3037-3046, 2000. PubMed: 11115848
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