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| 产品名称： | CAR47 |
|---|---|
| 商品货号： | TS197157 |
| Organism： | Homo sapiens, human |
| Tissue： | adrenal gland |
| Cell Type： | fibroblast |
| Product Format： | frozen |
| Morphology： | fibroblast-like |
| Culture Properties： | adherent |
| Biosafety Level： | 2 xa0Cells contain SV40 viral DNA sequences
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： | primary pigmented adrenocortical disease (PPNAD) and Carney complex |
| Age： | adult |
| Gender： | female |
| 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. |
| Storage Conditions： | liquid nitrogen vapor phase |
| Images： |  |
| 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. |
| Clinical Data： | adult female patient diagnosed with PPNAD, Carney complex, and Cushing syndrome |
| 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. |
| Complete Growth 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:
|
| Subculturing： | Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
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 |
| Cryopreservation： | Freeze Medium: Complete growth medium, 70%; FBS, 20%; DMSO, 10% Storage Temperature: Liquid nitrogen vapor phase |
| Culture Conditions： | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
| STR Profile： | 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 |
| Functional Tests： | validation of PRKA1A-inactivating mutation |
| Name of Depositor： | M. Nesterova |
| Year of Origin： | 2005 |
| References： | 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 |