vEPT (ATCC® CRL-2087)

Organism: Oryctolagus cuniculus, rabbit  /  Cell Type: Epithelial,fibroblast  / 

Organism Oryctolagus cuniculus, rabbit
Cell Type Epithelial,fibroblast
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
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.
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
Protocol: Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 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, which 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 with 5 to 10 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 xg for 5 to10 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 type I. 7. Place culture vessels in incubators at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Twice per week
Cryopreservation
Culture medium, 95%; DMSO, 5%
Name of Depositor U Hopfer
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

Basic Documentation
References

Romero MF, et al. Development and characterization of rabbit proximal tubular epithelial cell lines. Kidney Int. 42: 1130-1144, 1992. PubMed: 1280703