Clone C細胞
數量： 大量
運輸方式： 凍存運輸
生長狀態： 貼壁生長
細胞類型： 其他細胞類型
是否是腫瘤細胞： 0
物種來源： 其他
器官來源： 腎皮質
細胞形態： 上皮樣
ATCC Number： CRL-2531?
Designations: Clone C
Clone C細胞Depositors: Q Al-Awqati
Biosafety Level: 2 [Cells contain SV40 viral sequences ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Oryctolagus cuniculus deposited as rabbit
Morphology: epithelial
Source: Organ: kidney, cortex
Cell Type: intercalated; SV40 large T antigen transfected
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Comments: Clone C細胞Primary cultures of rabbit, bicarbonate-secreting, intercalated cells were isolated. [49845]
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. [49846]
Following transfection, a line of G418 resistant cells was obtained and designated IC250. These cells were subcloned (Clone C) and characterized. [49846]
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 oftrue bicarbonate-secreting intercalated epithelia cells. [49846]
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. [49846]
These cells reverse the polarity of several proteins and show different cytoskeltal organization depending on the seeding density. [49847]
The cell line may be used to study terminal differentiation of intercalated cells.
Propagation: ATCC 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
Clone C細胞15 ng/ml epidermal growth factor(do not filter)
25 ng/ml hydrocortisone
10% fetal bovine serum
Temperature: 32.0°C
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.
Preservation: culture medium 95%; DMSO, 5%
Related Products: recommended serum:ATCC 30-2020
References: 49845: van Adelsberg J, et al. Clone C細胞Isolation and culture of HCO3- -secreting intercalated cells. Am. J. Physiol. 256: C1004-C1011, 1989. PubMed: 2541617
49846: Edwards JC, et al. Conditional immortalization of bicarbonate-secreting intercalated cells from rabbit. Am. J. Physiol. 263: C521-C529, 1992. PubMed: 1325122
49847: 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
51589: 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
