UMB1949 (ATCC® CRL-4004)

Organism: Homo sapiens, human  /  Cell Type: Melanocytes immortalized with hTERT and SV40 large T antigen  /  Tissue: Kidney; angiomyolipoma  /  Disease: Tuberous sclerosis

Permits and Restrictions

View Permits View Restrictions

Organism Homo sapiens, human
Tissue Kidney; angiomyolipoma
Cell Type Melanocytes 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]
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.
Images
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.
  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 1.5 X 104 to 2.5 X 104 viable cells/cm2 is recommended.
  6. 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.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
  • License agreement required for commercial customer uses.
  • This material is distributed for research purposes only. A signed addendum to the ATCC Material Transfer Agreement must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
Restrictions

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 ATCC before shipment. The price listed above is for noncommercial and academic organizations only. Commercial and for-profit organizations should call for pricing.

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.