WT 9-7 (ATCC® CRL-2830)

Organism: Homo sapiens, human  /  Cell Type: epithelial  /  Tissue: kidney, cortex, proximal tubule, cyst  / 

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Organism Homo sapiens, human
Tissue kidney, cortex, proximal tubule, cyst
Cell Type epithelial
Product Format frozen
Morphology polygonal
Culture Properties adherent
Biosafety Level 2 [Cells contain SV-40 viral DNA sequences]
Age unknown
Gender female
Applications These new series of cyst-derived cell lines represent useful in vitro models for studying the cellular and molecular biology of ADPKD.
Storage Conditions liquid nitrogen vapor phase
Derivation

The WT 9-7 cell line was established in July, 1999 from a single cyst from a proximal cortical tubule taken from a patient with Autosomal dominant polycystic kidney disease (ADPKD).

WT 9-7 is one of a series of immortalized epithelial cell lines from over 30 individual renal cysts obtained from 11 patients with ADPKD.

The cells were transformed with a wild type recombinant adeno-SV40 virus.

Antigen Expression alanyl (membrane) aminopeptidase (ANPEP, CD13); Homo sapiens, expressed RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101 RefLoghman-Adham M, et al. An intrarenal renin-angiotensin system in autosomal dominant polycystic kidney disease. Am. J. Physiol. Renal Physiol. 287: F775-F788, 2004. PubMed: 15187005
large T antigen; Simian virus 40, expressed RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
Receptor Expression epidermal growth factor (EGF), expressed RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
Genes Expressed
angiotensinogen RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
polycystic kidney disease 1 RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
polycystic kidney disease 2 RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
Cellular Products
angiotensinogen RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
polycystic kidney disease 1 RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
polycystic kidney disease 2 RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
Comments These viruses contain an origin-defective (ori-) WT SV40 DNA cloned into the adenovirus vector in place of regions 1a and 1b. RefLoghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101
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 cmflask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: Flasks must be coated with Bovine Collagen type I solution (3.0 mg/mL) for 1 hour at 37°C
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 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 Bovine Collagen Type I (3 mg/mL) coated flasks.
    An inoculum of 5 x 103 to 1 x 104 viable cells/cm2 is recommended.
  6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 5 x 103 and 5 x 104 cells/cm2
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended
Medium Renewal: Two to three times weekly

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

Cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions

Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Note: Flasks must be coated with Bovine Collagen Type I solution (3.0 mg/mL) for 1 hour at 37°C.

Population Doubling Time 26 hours
Name of Depositor M Loghman-Adham
Year of Origin July, 1999
References

Loghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101

Loghman-Adham M, et al. An intrarenal renin-angiotensin system in autosomal dominant polycystic kidney disease. Am. J. Physiol. Renal Physiol. 287: F775-F788, 2004. PubMed: 15187005

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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.
Basic Documentation
References

Loghman-Adham M, et al. Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. Am. J. Physiol. Renal Physiol. 285: F397-F412, 2003. PubMed: 12734101

Loghman-Adham M, et al. An intrarenal renin-angiotensin system in autosomal dominant polycystic kidney disease. Am. J. Physiol. Renal Physiol. 287: F775-F788, 2004. PubMed: 15187005

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.