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SV7tert PDGF tumor-1 (ATCC® CRL-4008™)

Organism: Homo sapiens, human  /  Cell Type: Epithelial cells immortalized with hTERT and SV40 large T antigen expression  /  Tissue: Kidney; angiomyolipoma  / 

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Organism Homo sapiens, human
Tissue Kidney; angiomyolipoma
Cell Type Epithelial cells immortalized with hTERT and SV40 large T antigen expression
Product Format frozen
Morphology Epithelial-like
Culture Properties Adherent
Biosafety Level 2  [Cells 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.
Age 63 years
Gender female
Storage Conditions liquid nitrogen vapor phase
Karyotype This is a hypotetraploid cell line with many structural rearrangements, numerical losses and gains. The following eight derivatives were found to be present in low and high passage karyotypes: der(x)t(X;3)(q28;p21), der(1)t(1;17)(q10;p10), der(3)t(3;6)(p10:p10), i(8)(q10), i(12)(q10), der(13)t(13;21)(q10;q10), der(16)t(4;16)(q21;q24), add(20)(q13.3). Generally, the karyotyped passages contained the same complement of chromosome rearrangements, losses and gains.
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Derivation

CRL-4008 was derived from SV7tert (ATCC CRL-2461), a non-tumorigenic angiomyolipoma cell line immortalized with the SV40 large T antigen and human telomerase, by transduction with a retrovirus encoding PDGF-BB. 

The transduced cells were implanted into nude mice and formed tumors from which SV7tert PDGF tumor-1 (ATCC CRL-4008) was derived.The tumor-derived cells secrete over 18-fold more PDGF than pre-implantation cells, and demonstrate both autocrine transformation and epigenetic changes.
Antigen Expression This cell line is positive for the epithelial marker pan-cytokeratin as assessed by immunocytochemistry.
Cellular Products
Platelet Derived Growth Factor (PDGF-BB)
RefGovindarajan B, et al. Malignant transformation of human cells by constitutive expression of platelet-derived growth factor-BB. J. Biol. Chem. 280(14): 13936-13943, 2005. PubMed: 15695519
Tumorigenic Yes
Complete Growth 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%.
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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 1.0 to 2.0 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. An inoculum of 1.0 X 104 to 2.0 X 104 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C. Subculture when the cell concentration is between 8 X 104 to 1 X 105 cells/cm2.
Subcultivation ratio: A subcultivation ratio of 1:4 to 1:10 is recommended.
Medium renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X
CSF1PO: 10, 12
D13S317: 8
D16S539: 9, 11
D5S818: 11, 13
D7S820: 11
THO1: 6, 9.3
TPOX: 8, 11
vWA: 17, 18
Population Doubling Level (PDL) As part of our quality control, we have tested this cell line for its ability to grow for a minimum of 15 population doublings after recovery from cryopreservation. We have also compared its karyotype, telomerase expression level, growth rate, morphology and tissue-specific markers when first recovered from cryopreservation with that of cells at 10+ population doublings to ensure that there is no change in these parameters and that the cells are capable of extended proliferation.
Name of Depositor JL Arbiser
Year of Origin August 2003
References

Govindarajan B, et al. Malignant transformation of human cells by constitutive expression of platelet-derived growth factor-BB. J. Biol. Chem. 280(14): 13936-13943, 2005. PubMed: 15695519

Arbiser JL, et al. The generation and characterization of a cell line derived from a sporadic renal angiomyolipoma: use of telomerase to obtain stable populations of cells from benign neoplasms. Am. J. Pathol. 159: 483-491, 2001. PubMed: 11485907

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

Arbiser JL, et al. Functional tyrosine kinase inhibitor profiling: a generally applicable method points to a novel role of platelet-derived growth factor receptor-beta in tuberous sclerosis. Am. J. Pathol. 161(3): 781-786, 2002. PubMed: 12213705

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • This material is distributed for research purposes only. A signed hTERT addendum to the ATCC Material Transfer Agreement must be sent to ATCC in advance of shipment.
  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
SDS
SDS
Other Documentation
Other Document
Cell Micrograph
Restrictions

For commercial accounts, this cell line is only distributed under the terms of a fully signed and executed ATCC® Material Transfer Agreement and Addendum. If the commercial account is screening per completed Addendum, the recipient will be required to pay a Screening Fee (ATCC® ACS-2103™) of $4000.00.

Screening Use is defined as use of Biological Material in small molecule and biologic drug discovery, including initial target identification and validation, assay development, high throughput screening, hit identification, lead optimization, and selection of candidates for clinical development.

If the commercial account is not screening per the completed Addendum, the recipient will not be required to pay a Screening Fee.
References

Govindarajan B, et al. Malignant transformation of human cells by constitutive expression of platelet-derived growth factor-BB. J. Biol. Chem. 280(14): 13936-13943, 2005. PubMed: 15695519

Arbiser JL, et al. The generation and characterization of a cell line derived from a sporadic renal angiomyolipoma: use of telomerase to obtain stable populations of cells from benign neoplasms. Am. J. Pathol. 159: 483-491, 2001. PubMed: 11485907

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

Arbiser JL, et al. Functional tyrosine kinase inhibitor profiling: a generally applicable method points to a novel role of platelet-derived growth factor receptor-beta in tuberous sclerosis. Am. J. Pathol. 161(3): 781-786, 2002. PubMed: 12213705