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  /  Disease: Tuberous Sclerosis

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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]
Disease Tuberous Sclerosis
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
Gene products: Platelet Derived Growth Factor (PDGF-BB)
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

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This material requires that the Addendum for Commercial and For-Profit Organizations or the Addendum for Noncommercial and Academic Organizations be signed and returned to AT CC before shipment. The price listed above is for noncommercial and academic organizations only. Commercial and for-profit organizations should call for pricing.

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