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微信小章| 产品名称： | Clone C |
|---|---|
| 商品货号： | TS212338 |
| 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
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. |
| 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:
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| 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 |
| Deposited As： | rabbit |
| 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 |