DMEM HIGH GLUCOSE

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/ml), streptomycin (100 U/ml), 5 mM L-glutamine and 1.5 g/l sodium bicarbonate

Protocol tips
supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/ml), streptomycin (100 U/ml), 5 mM L-glutamine and 1.5 g/l sodium bicarbonate

Publication protocol

PAECs and PASMCs were cultured from porcine pulmonary arteries as previously described (18,21). PAECs and PASMCs, which were used between passages 2 and 6, were isolated from porcine pulmonary arteries obtained from a local abattoir within 1 h of slaughter. Endothelial cells were digested with 1.7 mg/ml collagenase II, then the pulmonary arteries were cut open along the longitudinal axis, and the residual endothelium was gently removed by scraping the luminal surface and washed away with phosphate-buffered saline (PBS). The adventitia layer was removed by blunt dissection, and the medial smooth muscle tissue was minced into 1 mm3 explant pieces. PAECs and the explant pieces of smooth muscle were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/ml), streptomycin (100 U/ml), 5 mM L-glutamine and 1.5 g/l sodium bicarbonate. The medium was changed every 2 days. The cells were cultured in an incubator at 37ºC and maintained in a humidified atmosphere with 5% CO2. The purity and identity of the endothelial cells and smooth muscle cells were verified by their typical morphological patterns and by immunofluorescent staining for factor VIII-related antigen and α-smooth muscle actin (α-SMA), respectively (16,21). All passages were performed using 0.05% trypsin and 0.02% EDTA. Cell viability was assessed using the CCK-8 assay.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Mammalian cell culture media PPAEC using DMEM HIGH GLUCOSE from EuroClone Online Store.

Manufacturer protocol

Download the product protocol from EuroClone Online Store for DMEM HIGH GLUCOSE below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Mammalian cell culture media PPAEC using DMEM HIGH GLUCOSE from EuroClone Online Store. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.