Clone C (ATCC® CRL-2531)

Organism: Oryctolagus cuniculus, rabbit  /  Cell Type: intercalated; SV40 large T antigen transfecte  / 

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Organism Oryctolagus cuniculus, rabbit
Cell Type intercalated; SV40 large T antigen transfecte
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain SV40 viral sequences
Applications
Cells were transfected by electroporation with the pZipSVtsA58 plasmid encoding a temperature-sensitive large T antigen of SV40 plus the neomycin resistance gene under the control of an SV40 promoter.
They express appropriate ultrastructural features, bind peanut lectin in an apical pattern, are rich in carbonic anhydrase and stain for proton-adenosinetriphosphatase in a basolateral pattern.
The cell line may be used to study terminal differentiation of intercalated cells.
Derivation
Primary cultures of rabbit, bicarbonate-secreting, intercalated cells were isolated.
Following transfection, a line of G418 resistant cells was obtained and designated IC250. These cells were subcloned (Clone C) and characterized.
Comments
Primary cultures of rabbit, bicarbonate-secreting, intercalated cells were isolated.
Cells were transfected by electroporation with the pZipSVtsA58 plasmid encoding a temperature-sensitive large T antigen of SV40 plus the neomycin resistance gene under the control of an SV40 promoter.
Following transfection, a line of G418 resistant cells was obtained and designated IC250. These cells were subcloned (Clone C) and characterized.
The cells divide continuously at the permissive temperature of 32C. At the restrictive temperature of 40C, they cease dividing and assume morphological and transport properties of true bicarbonate-secreting intercalated epithelia cells.
They express appropriate ultrastructural features, bind peanut lectin in an apical pattern, are rich in carbonic anhydrase and stain for proton-adenosinetriphosphatase in a basolateral pattern.
These cells reverse the polarity of several proteins and show different cytoskeltal organization depending on the seeding density.
The cell line may be used to study terminal differentiation of intercalated cells.
Complete Growth Medium The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No. 30-2006.To make the complete growth medium, add the following components to the base medium:
  • 0.005 mg/ml insulin
  • 0.005 mg/ml transferrin
  • 1.75 ng/ml selenious acid
  • 15 ng/ml epidermal growth factor(do not filter)
  • 25 ng/ml hydrocortisone
  • 10% fetal bovine serum

Subculturing
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature until the cells detach.
Add fresh culture medium, aspirate and dispense into new culture flasks.
Cryopreservation
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 32.0°C
Name of Depositor Q Al-Awqati
References

van Adelsberg J, et al. Isolation and culture of HCO3- -secreting intercalated cells. Am. J. Physiol. 256: C1004-C1011, 1989. PubMed: 2541617

Edwards JC, et al. Conditional immortalization of bicarbonate-secreting intercalated cells from rabbit. Am. J. Physiol. 263: C521-C529, 1992. PubMed: 1325122

van Adelsberg J, et al. An induced extracellular matrix protein reverses the polarity of band 3 in intercalated epithelial cells. Cell 76: 1053-1061, 1994. PubMed: 8137422

Vijayakumar S, et al. Hensin remodels the apical cytoskeleton and induces columnarization of intercalated epithelial cells: processes that resemble terminal differentiation. J. Cell Biol. 144: 1057-1067, 1999. PubMed: 10085301

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

van Adelsberg J, et al. Isolation and culture of HCO3- -secreting intercalated cells. Am. J. Physiol. 256: C1004-C1011, 1989. PubMed: 2541617

Edwards JC, et al. Conditional immortalization of bicarbonate-secreting intercalated cells from rabbit. Am. J. Physiol. 263: C521-C529, 1992. PubMed: 1325122

van Adelsberg J, et al. An induced extracellular matrix protein reverses the polarity of band 3 in intercalated epithelial cells. Cell 76: 1053-1061, 1994. PubMed: 8137422

Vijayakumar S, et al. Hensin remodels the apical cytoskeleton and induces columnarization of intercalated epithelial cells: processes that resemble terminal differentiation. J. Cell Biol. 144: 1057-1067, 1999. PubMed: 10085301