Primary mouse lung endothelial cells were derived and immortalized with polyoma middle T (PmT) virus to generate the Immortalized Mouse Lung Endothelial Cells – PmT. The cells are valuable for downstream applications to study the lung endothelium, as they can continuously divide in culture. It may be useful for studies relating to diseases and understanding the biology of lung endothelial cells.Immortalization Method: Immortalized by infecting with polyoma middle T (PmT) virusBioSafety Level: IISpecies: MouseSource Organ: LungGrowth Properties: AdherentMorphology: EndothelialPopulation Doubling: N/ASeeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.Markers: N/ADonor Age: N/ADonor Gender: N/ADonor Ethnicity: N/APropagation: This cell line requires coated cell culture vessels of collagen (final concentration of 30ug/ml; G422) and fibronectin (final concentration of 10ug/ml; TM059) in 0.1% gelatin (TM063). The base medium for this cell line is a 1:1 (recommend 400ml:400ml) ratio mix of Prigrow VI and Prigrow IV Series medium available at abm, Cat. No. TM016, and Cat. No. TM004 respectively. To make the completed growth medium, add the following components to the base medium: 100 mg heparin (Sigma; H3142) dissolved in 10 ml of Prigrow IV Medium (TM004), 10 ml Penicillin/Streptomycin Solution (G255), and 20 ml L-glutamine (G275). Filter the medium with 0.2 uM filter before adding fetal bovine serum (TM999)* which is first heat-inactivated before use to final concentration of 10% and 50 mg of endothelial mitogen (Endothelial Cell Growth Supplement) (Generon). Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
Recommended split ratio of 1:4 or 1:8. Preform a PBS wash before using 0.25% Trypsin-EDTA (TM050) to subculture.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.Quality Control: N/AShipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C