UMB1949細(xì)胞

數(shù)量： 大量

運(yùn)輸方式： 凍存運(yùn)輸

是否是腫瘤細(xì)胞： 0

物種來源： 人

年限： 36 years

細(xì)胞形態(tài)： 上皮樣

組織來源： angiomyolipoma

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

ATCC Number： CRL-4004?

相關(guān)**： 其他**

器官來源： 腎臟

Designations: UMB1949

Depositors: JL Arbiser

Biosafety Level: 2 UMB1949細(xì)胞[cells contain 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

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Isolation: Isolation date: June 2004

Applications: UMB1949細(xì)胞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.

As such, this cell line can be used to study signal transduction and drug efficiency in tuberous sclerosis.

The UMB1949 cell line was established by sequential introduction of the SV40 large T antigen and human telomerase into human angiomyolipoma cells.

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

DNA Profile (STR): Amelogenin: XY

CSF1PO: 10, 11

D13S317: 12, 13

D16S539: 12

D5S818: 11

D7S820: 9, 10

THO1: 6, 8

TPOX: 12

vWA: 18

Cytogenetic Analysis: UMB1949細(xì)胞This cell line is of male origin and 1/2 to 2/3 of the total cell population is pseudodiploid, the rest of the cells fall in the tetraploid range. Consistent cytogenetic changes include chromosome 10 and 19 aberration, and chromosome 4 monosomy. Some cells showed loss of the Y chromosome and many of the examined cells contained random chromosomal aberrations.

Age: 36 years

Gender: male

Ethnicity: Caucasian

Comments: The UMB1949 cell line was established by sequential introduction of the SV40 large T antigen and human telomerase into human angiomyolipoma cells. Angiomyolipomas are benign tumors of the kidney which originate from putative perivascular epithelioid cells that may undergo differentiation into cells with features of melanocytes, smooth muscle or fat cells. UMB1949 cells have a defined 5bp deletion in exon 33 of tuberin (Tsc2) and mutations in tuberin (and/or hamartin) cause tuberous sclerosis. As such, this cell line can be used to study signal transduction and drug efficiency in tuberous sclerosis. [16173554]

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: UMB1949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 1.5 X 104 to 2.5 X 104 viable cells/cm2 is recommended.

Incubate cultures at 37.0°C. Subculture when the cell concentration is between 5 X 104 to 7 X 104 cells/cm2.

Subcultivation ratio: A subcultivation ratio of 1:3 to 1:5 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: about 29 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

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

16173554: Lim SD , et al. Expression of the neural stem cell markers NG2 and L1 in human angiomyolipoma: are angiomyolipomas neoplasms of stem cells? Mol. Med. 13(3-4): 160-165, 2007. PubMed: 17592550