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Immortalized-Mouse-Kupffer-Cells-(ImKC)

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Immortalized-Mouse-Kupffer-Cells-(ImKC)

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	Print Version
BioSafety Level	II
Organism	H-2K^b-tsA58 transgenic mice
Source Organ	Liver
Donor Gender	Male
Growth Properties	Adherent
Morphology	Macrophage-like
Population Doubling	~ 24 hrs
Recommended Seeding Density	4000-8000 cells/cm²
Markers	F4/80, CD11b, CD14, and TLR4
Applications	For Research Use Only
Immortalization Method	Isolated from transgenic mice expressing thermolabile mutant tsA58 of SV40 40 large T antigen
Description	Immortalized Mouse Kupffer Cells (ImKC) are isolated from male H-2K^b-tsA58 transgenic mice which expresses the thermolabile mutant tsA58 of SV40 40 large T antigen and were sorted through identifying the F4/80 marker. The immortalized cells expresses p53 levels with increases telomerase activity and retains the some functional features compared to the primary cells. ImKC cells can propagate without the presence of IFNy at 37°C, and are able to produce proinflammatory cytokines when induced with lipopolysaccharide (LPS). These cells are valuable for studies in proinflammatory cytokines (TNF-α, IL-6 and IL-1β) and their roles in Kuppfer cells, in processes in liver injury, and for research in chronic diseases.
Procedure Overview
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Propagation	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 II medium available from ABM (TM002). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999) which is first heat-inactivated before use to final concentration of 10% and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. To induce cytokine production stimulate cells with 100 ng/ml LPS for 1 hour.
Quality Control	1) Flow cytometry utilized to analyze cell surface markers F4/80, CD11b, CD14, and TLR4; 2) Western blot analysis on p53 expression; 3) qRT-PCR used to analyze level of LPS induced cytokine production
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. 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 strongly 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.