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Aortic endothelial cells cultured in matrix (Shire)

Porcine aortic endothelial cells (PAEs) are isolated from the aorta of healthy pigs by collagenase digestion, and cultured on gelatin particles. 

Cells are maintained at 37°C in a humidified 5% CO2/95% air incubator and cultured up to passage 6 in DMEM supplemented with penicillin (100 U/mL), streptomycin (100 μg/mL), glutamine (2.0 mmol/L), and calf serum (10%). Gelfoam is cut into 2.5×1.0×0.3-cm blocks. The Gelfoam blocks are hydrated in PBS and placed in 35×10-mm tissue culture dishes; 0.1 mL of a 0.8×106 cells/mL PAE cell suspension is placed directly on the sponge, thereby seeding 0.8×105 cells/block. The cells are placed at 37°C in a humidified 5% CO2/95% incubator and allowed to adhere for 2 hours. The cell-loaded Gelfoam blocks are subsequently placed in 17×100-mm polypropylene tubes containing 2 mL of medium and incubated for up to 2 weeks at a 45° angle. Growth medium is changed every 72 hours.

In some of the applications 4.0 × 1.0 × 0.3 cm3 blocks of sterile Gelfoam® were seeded with 1.25 –1.66 × 105cells/cm3 of Gelfoam® and incubated for ≈ 2 weeks until confluent in Endothelial Basal Medium supplemented with FBS (5%) and gentamicin (50 μg/mL).

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Aortic endothelial cells cultured in matrix