WT 9-12 (ATCC® CRL-2833)

Organism: Homo sapiens, human  /  Cell Type: epithelial  / 

Organism Homo sapiens, human
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
WT 9-12 is one of a series of immortalized epithelial cell lines from over 30 individual renal cysts obtained from 11 patients with ADPKD. These new series of cyst-derived cell lines represent useful in vitro models for studying the cellular and molecular biology of ADPKD. Antigen: SV40 large T antigen, expressed [PubMed: 12734101]
The WT 9-12 cell line was established in July, 1999 from a single cyst from a cortical tubule taken from a patient with autosomal dominant polycystic kidney disease (ADPKD).
Storage Conditions liquid nitrogen vapor phase
Derivation
The WT 9-12 cell line was established in July, 1999 from a single cyst from a cortical tubule taken from a patient with autosomal dominant polycystic kidney disease (ADPKD). The cells have both distal and proximal characteristics. They can be subcloned to obtain clones with proximal or distal markers. The cells were transformed with a wild type recombinant adeno-SV40 virus. These viruses contain an origin-deficient (ori-) WT SV40 DNA cloned into the adenovirus vector in place of regions 1a and 1b [PubMed: 12734101]. WT 9-12 is one of a series of immortalized epithelial cell lines from over 30 individual renal cysts obtained from 11 patients with ADPKD. These new series of cyst-derived cell lines represent useful in vitro models for studying the cellular and molecular biology of ADPKD. Antigen: SV40 large T antigen, expressed [PubMed: 12734101]
Clinical Data
The WT 9-12 cell line was established in July, 1999 from a single cyst from a cortical tubule taken from a patient with autosomal dominant polycystic kidney disease (ADPKD).
female
Antigen Expression
alanyl (membrane) aminopeptidase (ANPEP, CD13); Homo sapiens, expressed
Receptor Expression
epidermal growth factor (EGF), expressed
Genes Expressed
epithelial sodium channel, ENaC,alanyl (membrane) aminopeptidase (ANPEP, CD13); Homo sapiens, expressed
Comments
The WT 9-12 cell line was established in July, 1999 from a single cyst from a cortical tubule taken from a patient with autosomal dominant polycystic kidney disease (ADPKD). The cells have both distal and proximal characteristics. They can be subcloned to obtain clones with proximal or distal markers. The cells were transformed with a wild type recombinant adeno-SV40 virus. These viruses contain an origin-deficient (ori-) WT SV40 DNA cloned into the adenovirus vector in place of regions 1a and 1b [PubMed: 12734101]. WT 9-12 is one of a series of immortalized epithelial cell lines from over 30 individual renal cysts obtained from 11 patients with ADPKD. These new series of cyst-derived cell lines represent useful in vitro models for studying the cellular and molecular biology of ADPKD.
Antigen: SV40 large T antigen, expressed [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
Protocol:
  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 37C 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 3 to 5 X 10(3) viable cells/sq. cm is recommended.
  6. Incubate cultures at 37C.
Interval: Maintain cultures at a cell concentration between 5 X 10(3) and 8 X 10(4) cells/sq. cm
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Two to three times weekly
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Growth Conditions: Flasks must be coated with Bovine Collagen type I solution (3.0 mg/ml) for 1 hour at 37C.
Population Doubling Time 38 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

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