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生長狀態(tài)：貼壁生長
數(shù)量：大量
細胞類型：上皮細胞
ATCC Number：CRL-2833?
相關疾病：autosomal dominant polycystic kidney disease
是否是腫瘤細胞：0
物種來源：人
年限：unknown
器官來源：腎臟
運輸方式：凍存運輸
細胞形態(tài)：多邊形
組織來源：cyst from a distal and proximal cortical tubule
規(guī)格：0.2ml Designations: WT 9-12
Depositors: ?M Loghman-Adham
Biosafety Level:2 [Cells contain SV-40 viral DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Homo sapiens
Morphology:polygonal


Source: Organ: kidney
Tissue: cyst from a distal and proximal cortical tubule
Disease: autosomal dominant polycystic kidney disease (ADPKD) [89940 ]
Cell Type: epithelial
Cellular Products:epithelial sodium channel, ENaC
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: Isolation date: July, 1999
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).
Receptors:epidermal growth factor (EGF), expressed [89940 ]
Antigen Expression:alanyl (membrane) aminopeptidase (ANPEP, CD13); Homo sapiens, expressed [89940 ]
Age: unknown
Gender: female
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]
Propagation: ATCC 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%.
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.
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
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 38 hours
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
derived from same individual:ATCC CRL-2830
derived from same individual:ATCC CRL-2832
derived from same individual:ATCC CRL-2831
References: 89940: 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