Immortalized Mouse Lung Clara Cells - Conditionally (C22)

Cat. No.
T0092
Unit
1x106 cells / 1.0 ml
Price
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Cat. No. T0092
Name Immortalized Mouse Lung Clara Cells - Conditionally (C22)
Description

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.

Presence of CC10, SP-A, SP-B and absence of SP-C confirmed by RT-PCR.

Organism Mouse (M. musculus)
Tissue Lung
Donor History H-2Kb-tsA58 transgenic mouse
Growth Properties

Adherent, spindle-shaped

Cell Type Immortalized Cells
Unit 1x106 cells / 1.0 ml
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions

Ship with dry ice.

Product Format Frozen
Intended Use This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions

PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. PriGrow III (TM003+ 10% FBS + 100 U/ml rmIFNG (Z200085) + 5 ug/ml transferrin + 25 ng/ml rmEGF (Z200025) + 10 ug/ml insulin (Z101065) + 7.5 ug/ml ECGS (TM141+ 0.25 ug/ml endothelin-1 + 20 ng/ml T3 + 0.36 ug/ml hydrocortisone + 1% Penicillin/Streptomycin Solution (G255), 33.0°C, 5% CO₂.

Please note: the optimal incubator temperature for cell proliferation is 33.0°C.

For information regarding cell differentiation, please refer to the "Differentiation Protocol" PDF under the Documents Tab.

Unpacking and Storage Instructions

1. Visually examine the packaging containers for signs of leakage or breakage.

2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.

To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.

 

Thawing Protocol

1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.

2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.

3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.

4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.

5. Incubate the cells at the recommended conditions.

Subculture Protocol

Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.

1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.

2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 33°C, for several minutes to facilitate detachment.

3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.

4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.

5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.

6. Incubate the cells at the recommended conditions.

Cryopreservation

Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.

Immortalization Method

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

Expression

CC10, SP-A, SP-B, SP-D, and absence of SP-C

STR Profiling

4-2 : 18.3,19.3

5-5 : 13,

6-4 : 18,

6-7 : 12,17

9-2 : 15,

12-1 : 16,

15-3 : 26.3,

18-3 : 16,

X-1 : 25,

Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0092.
Warranty abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period”.
Disclaimer
1. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.

2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).

3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.

4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.

5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Depositor Saint Louis University
Application

Research Use Only.

Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0092
Print & Download Datasheet
  • Demello, D. E., Mahmoud, S., Ryerse, J., & Hoffmann, J. W. (2002). Generation and characterization of a conditionally immortalized lung clara cell line from the H-2Kb-tsA58 transgenic mouse. In Vitro Cellular & Developmental Biology-Animal, 38(3), 154-164.

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