Clone C細胞

數(shù)量： 大量

運輸方式： 凍存運輸

生長狀態(tài)： 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(zhì)

細胞形態(tài)： 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