UMB1949 永生化细胞-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心CRL-4004

UMB1949 永生化细胞-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心CRL-4004
Cat Number: CRL-4004
种属来源Organism: Homo sapiens, human
细胞类型Cell Type: Melanocyte
疾病类型：Disease Tuberous Sclerosis
供应厂家Supplier: BioVector NTCC Inc.    

http://www.biovector.net

Organism

Homo sapiens, human

TissueKidney; angiomyolipoma

Cell TypeMelanocyte immortalized with hTERT and SV40 large T antigen

Product Format  frozen

MorphologyEpithelial-like

Culture Properties  Adherent

Biosafety Level2  [Cells contain SV40 viral DNA sequences]

DiseaseTuberous sclerosis

Age36 years

GenderMale

Applications

This cell line can be used to study signal transduction and drug efficiency in tuberous sclerosis.KaryotypeThis 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.Images DerivationThe UMB1949 cell line was established by sequential introduction of the SV40 large T antigen and human telomerase into human angiomyolipoma cells. Antigen ExpressionAntigen expression: positive for epithelial marker pan-cytokeratin (immunocytochemistry)CommentsAngiomyolipomas 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 MediumThe base medium for this cell line is -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%. SubculturingVolumes 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.CryopreservationFreeze medium: complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phaseCulture ConditionsTemperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%STR ProfileAmelogenin: XY
CSF1PO: 10, 11
D13S317: 12, 13
D16S539: 12
D5S818: 11
D7S820: 9, 10
THO1: 6, 8
TPOX: 12
vWA: 18Population 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 DepositorJL ArbiserYear of OriginJune 2004ReferencesArbiser 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