產(chǎn)品名稱 |
WT 9-7 |
商品貨號(hào) |
B208743 |
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]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
Disease |
autosomal dominant polycystic kidney disease (ADPKD) |
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 Ref 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 Ref 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
large T antigen; Simian virus 40, expressed Ref 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 |
Receptor Expression |
epidermal growth factor (EGF), expressed Ref 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 |
Genes Expressed |
|
Cellular Products |
angiotensinogen Ref  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
polycystic kidney disease 1 Ref  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
polycystic kidney disease 2 Ref  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 |
Comments |
These viruses contain an origin-defective (ori-) WT SV40 DNA cloned into the adenovirus vector in place of regions 1a and 1b. Ref 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 |
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 flask; 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 (supplied stock concentration is 3.0 mg/mL; PureCol® Advanced Biomatrix Catalog No. 5005-B) for 1 hour at 37°C
- Remove and discard culture medium.
-
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.
-
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.
-
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 Bovine Collagen Type I coated flasks.
An inoculum of 5 x 103 to 1 x 104 viable cells/cm2 is recommended.
- 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% |
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.
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