人妻少妇偷人精品无码丨色婷婷av久久久久久久丨欧美xxxx做受性欧美88丨欧洲女人牲交视频免费丨亚洲精品久久久av无码专区

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.

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 coated flasks.
   An inoculum of 5 x 10³ to 1 x 10⁴ viable cells/cm² is recommended.
6. Incubate cultures at 37°C.

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.

Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO₂), 5%

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.