| 產品名稱 |
vEPT |
| 商品貨號 |
B165923 |
| Organism |
Oryctolagus cuniculus, rabbit |
| Tissue |
kidney; proximal tubule |
| Cell Type |
Epithelial,fibroblast |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
1
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 |
normal |
| Age |
2 kg |
| Gender |
male |
| Applications |
Currently bovine collagen type I is used as a matrix. However, SV40 T antigen expression was not essential for immortalization, since neither SV40 T antigen nor G418 resistance was detected in the vEPT cell line. They retain electrolyte transport characteristics of the proximal tubule and receptor and signaling mechanisms for angiotensin II. They express cytokeratins 8, 10+11, and 19. The vEPT cell line was derived in 1989 from primary cultures of proximal tubules (S1 segment) microdissected from superficial slices of cortex from the kidney of a normal 2-kg male New Zealand rabbit by M. Romero and U. Hopfer. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
The vEPT cell line was derived in 1989 from primary cultures of proximal tubules (S1 segment) microdissected from superficial slices of cortex from the kidney of a normal 2-kg male New Zealand rabbit by M. Romero and U. Hopfer. The cells were originally grown on porous inorganic filters coated with poly-L-lysine. Currently bovine collagen type I is used as a matrix. After several passages the cells were co-cultured with irradiated fibroblasts producing a recombinant retrovirus encoding SV40 large T antigen and G418 resistance. However, SV40 T antigen expression was not essential for immortalization, since neither SV40 T antigen nor G418 resistance was detected in the vEPT cell line. These cells display properties of epithelial cells, forming confluent monolayers with apical microvilli, tight junctions, and convolutions of the basolateral plasma membrane. They retain electrolyte transport characteristics of the proximal tubule and receptor and signaling mechanisms for angiotensin II. They express cytokeratins 8, 10+11, and 19. |
| Clinical Data |
The vEPT cell line was derived in 1989 from primary cultures of proximal tubules (S1 segment) microdissected from superficial slices of cortex from the kidney of a normal 2-kg male New Zealand rabbit by M. Romero and U. Hopfer. male |
| Genes Expressed |
cytokeratins 8, 10 + 11, and 19 |
| Cellular Products |
cytokeratins 8, 10 + 11, and 19 |
| Comments |
The vEPT cell line was derived in 1989 from primary cultures of proximal tubules (S1 segment) microdissected from superficial slices of cortex from the kidney of a normal 2-kg male New Zealand rabbit by M. Romero and U. Hopfer. The cells were originally grown on porous inorganic filters coated with poly-L-lysine. Currently bovine collagen type I is used as a matrix. After several passages the cells were co-cultured with irradiated fibroblasts producing a recombinant retrovirus encoding SV40 large T antigen and G418 resistance. However, SV40 T antigen expression was not essential for immortalization, since neither SV40 T antigen nor G418 resistance was detected in the vEPT cell line. These cells display properties of epithelial cells, forming confluent monolayers with apical microvilli, tight junctions, and convolutions of the basolateral plasma membrane. They retain electrolyte transport characteristics of the proximal tubule and receptor and signaling mechanisms for angiotensin II. They express cytokeratins 8, 10+11, and 19. |
| Complete Growth Medium |
A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12K medium containing 1.2 g/L sodium bicarbonate, 50 nM hydrocortisone and 15 mM HEPES, 95%; fetal bovine serum, 5%
|
| 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.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which 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.
- To remove trypsin-EDTA solution, transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
- Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels that have been precoated with a thin film of Bovine Collagen I.
-
Place culture vessels in incubators at 37°C.
Subcultivation Ratio: 1:2 to 1:3
Medium Renewal: Every 2 to 3 days
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 described above 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% |
| Name of Depositor |
U Hopfer |
| Deposited As |
Oryctolagus cuniculus |
| Passage History |
The vEPT cell line was derived in 1989 from primary cultures of proximal tubules (S1 segment) microdissected from superficial slices of cortex from the kidney of a normal 2-kg male New Zealand rabbit by M. Romero and U. Hopfer. The cells were originally grown on porous inorganic filters coated with poly-L-lysine. Currently bovine collagen type I is used as a matrix. After several passages the cells were co-cultured with irradiated fibroblasts producing a recombinant retrovirus encoding SV40 large T antigen and G418 resistance. However, SV40 T antigen expression was not essential for immortalization, since neither SV40 T antigen nor G418 resistance was detected in the vEPT cell line. These cells display properties of epithelial cells, forming confluent monolayers with apical microvilli, tight junctions, and convolutions of the basolateral plasma membrane. They retain electrolyte transport characteristics of the proximal tubule and receptor and signaling mechanisms for angiotensin II. They express cytokeratins 8, 10+11, and 19. |
| Year of Origin |
1989 |
| References |
Romero MF, et al. Development and characterization of rabbit proximal tubular epithelial cell lines. Kidney Int. 42: 1130-1144, 1992. PubMed: 1280703
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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