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UMB1949

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價格:詢價

編號:B175505

品牌:Mingzhoubio

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DNA

RNA

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產品名稱

UMB1949

商品貨號

B175505

Organism

Homo sapiens, human

Tissue

Kidney; angiomyolipoma

培養基

Cell Type

Melanocyte immortalized with hTERT and SV40 large T antigen

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.

Disease

Tuberous sclerosis

Age

36 years

Gender

Male

Applications

This cell line can be used to study signal transduction and drug efficiency in tuberous sclerosis.

Storage Conditions

Liquid nitrogen vapor phase

Karyotype

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. UMB1949 cells have a defined 5bp deletion in exon 33 of tuberin (Tsc2) and mutations in tuberin (and/or hamartin) cause tuberous sclerosis.

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Derivation

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

Antigen Expression

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

Comments

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.

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.

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: 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.
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°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

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Aminal Cells: A Manual of Basic Techniques by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.

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: XY

CSF1PO: 10, 11

D13S317: 12, 13

D16S539: 12

D5S818: 11

D7S820: 9, 10

THO1: 6, 8

TPOX: 12

vWA: 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

June 2004

References

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

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

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

Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13.

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