Immortalized Mouse Lung Clara Cells-Conditionally (C22)

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.

Immortalization Method: Isolated from transgenic mouse carrying a temperature-sensitive simian virus 40 tumor antigen (tsA58)

BioSafety Level: II

Species: Mouse

Source Organ: Lung

Growth Properties: Adherent

Morphology: Spindle shaped

Population Doubling: N/A

Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.

Markers: 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

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow III medium available at abm (TM003). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, 100 U/ml γ-IFN (Z200085), 5 µg/ml transferrin, 0.025 µg/ml EGF(Z200025), 10 µg/ml insulin(Z101065), 7.5 µg/ml ECGS(TM141), 0.25 µg/ml endothelin-1, 20 ng/ml T3 , 0.36 µg/ml hydrocortisone and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. To promote cell differentiation, withdraw γ-IFN from the culture medium, and incubate the cells at 39°C for 24-48 hours.
Carbon dioxide (CO₂): 5%, Temperature: 33.0°C. * abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: N/A

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C