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Print Version
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BioSafety Level
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II
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Organism
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H-2Kb-tsA58 transgenic mouse
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Source Organ
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Lung
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Growth Properties
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Adherent
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Morphology
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Under proliferation phase: cell size varies from 10 to 25 μm. Smaller cells are spindle shaped with clear cytoplasma and grow at a slower rate than larger cells, which are rounded in shape with a vacuolated cytoplasma and a faster doubling time.
Upon differentiation: cells increase in diameter and acquire clear to foamy vacuoles.
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Recommended Seeding Density
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Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
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Markers
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1) Presence of CC10, SP-A, SP-B and absence of SP-C confirmed by RT-PCR; 2) Synthesis and secretion of CC10, SP-A, SP-B and SP-D proteins confirmed by immunoblotting
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Applications
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For Research Use Only
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Immortalization Method
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Isolated from transgenic mouse carrying a temperature-sensitive simian virus 40 tumor antigen (tsA58)
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Description
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Clara cells play a role in the maintenance of normal bronchiolar epithelium by secreting Clara Cell Secretory Proteins (i.e. CC10) to protect the respiratory tract against oxidative stress and inflammation. The C22 is a conditionally immortalized mouse lung clara cells isolated from the mouse harboring thermolabile mutation (tsA58) under the control of an interferon (IFN)-γ-inducible H-2Kb promoter and a temperature-sensitive simian virus 40 large T antigen. The functional expression of the SV40 large T antigen is induced by culturing the cells in vitro in medium containing IFN γ at a temperature permissive (33°C). At a non-permissive temperature (37°C-39°C), the cells cease to proliferate.
C22 resembles closely to primary clara cells and constitutively expresses and secretes surfactant proteins SP-A, SP-B, SP-D and CC10. This cell line is useful in studying lung epithelial physiology, as well as studying the role of CC10 in pulmonary inflammation and viral infection.
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Procedure Overview
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Propagation
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Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow III medium available from ABM (TM003). To make the complete growth medium, add the following components to the base medium: 10% fetal bovine serum (TM999), 100 U/ml ?-IFN, 5 μg/ml transferrin, 0.025 μg/ml EGF, 10 μg/ml insulin, 7.5 μg/ml ECGS, 0.25 μg/ml endothelin-1, 20 ng/ml T3 , 0.36 μg/ml hydrocortisone and Penicillin/Streptomycin(G255). Atmosphere: air: 95%, CO2: 5%; Temperature: 33.0°C. To promote cell differentiation, withdraw ?-IFN from the culture medium, and incubate the cells at 39°C for 24-48 hours.
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Reference
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1) deMello, D. E., et al (2002). Generation and Characterization of a Conditionally Immortalized Lung Clara Cell Line from the H-2Kb-tsA58 Transgenic Mouse. In Vitro Cell Dev Biol Anim. 38 (3):154-64.
2) Elizur, A., et al (2007). Clara Cells Impact the Pulmonary Innate Immune Response to LPS. Am J Physiol Lung Cell Mol Physiol. 293(2):L383-92.
3) Elizur, A., et al (2008). Tumor Necrosis Factor-alpha from Macrophages Enhances LPS-induced Clara Cell Expression of Keratinocyte-derived Chemokine. Am J Respir Cell Mol Biol.38(1):8-15.
4) Atkinson, J. J., et al (2008). Clara Cell Adhesion and Migration to Extracellular Matrix. Respir Res. 7(9):1.
5) Adair-Kirk, T.L., et al (2008). Distal Airways in Mice Exposed to Cigarette Smoke: Nrf2-Regulated Genes are Increased in Clara Cells. Am J Respir Cell Mol Biol. 39(4):400-11.
6) Betsuyaku, T., et al (2013). Bronchiolar Epithelial Catalase is Diminished in Smokers with Mild COPD. Eur Respir J42(1):42-53.
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Disclaimer
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聯(lián)系方式
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何經(jīng)理，合肥星肽生物科技有限公司，Tel：0551-63802898 400-8702-898，
E-mail：[email protected] http://m.cnqiepian.com QQ:514713116
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