SV7tert PDGF tumor-1細(xì)胞

運(yùn)輸方式： 凍存運(yùn)輸

組織來源： angiomyolipoma

數(shù)量： 大量

生長狀態(tài)： 貼壁生長

是否是腫瘤細(xì)胞： 0

物種來源： 人

細(xì)胞形態(tài)： 上皮樣

器官來源： 腎臟

年限： 63 years

ATCC Number： CRL-4008?

相關(guān)**： 其他**

Designations: SV7tert PDGF tumor-1

Depositors: JL Arbiser

SV7tert PDGF tumor-1細(xì)胞Biosafety Level: 2 [cells containing SV40 viral DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial-like

Source: Organ: kidney

Tissue: angiomyolipoma

Disease: tuberous sclerosis

Immortalization Method: introduction of telomerase into SV40-transfected angiomyolipoma cells

Cellular Products: Platelet Derived Growth Factor (PDGF-BB) [16173555]

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Isolation: Isolation date: August 2003

Applications: In addition, it has been verified that no gross changes are observed in karyotype and morphology during the first 10 population doublings.

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.

The tumor-derived cells secrete over 18-fold more PDGF than pre-implantation cells, and demonstrate both autocrine transformation and epigenetic changes.

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.

Tumorigenic: YES

Antigen Expression: Positive for epithelial marker pan-cytokeratin (immunocytochemistry)

DNA Profile (STR): 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

Cytogenetic Analysis: SV7tert PDGF tumor-1細(xì)胞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.

Age: 63 years

Gender: female

Comments: 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. [16173555]

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. In addition, it has been verified that no gross changes are observed in karyotype and morphology during the first 10 population doublings.

Propagation: ATCC 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%.

Temperature: 37.0°C

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Subculturing: Protocol: 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 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.

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: SV7tert PDGF tumor-1細(xì)胞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.

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.

Incubate cultures at 37.0°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

Preservation: Freeze medium: complete growth medium, 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Doubling Time: approximately 28 hours

Related Products: Recommended medium (without the additional serum described under ATCC Medium): ATCC 30-2002

Recommended serum: ATCC 30-2020

0.25% (w/v) Trypsin - 0.53mM EDTA in Hank's BSS (w/o Ca++, Mg ++): ATCC 30-2101

Cell culture tested DMSO: ATCC 4-X

Phosphate-buffered saline: ATCC 30-2200

Parental cell line: ATCC CRL-2461

References: 47354: Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

53507: 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

16173555: 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

16173562: 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