Isolation and culture of endothelial cells from the mesenteric vascular bed

Methods are described for the enzymatic isolation of endothelial cells from rat and rabbit mesenteric arteries and veins. The mesenteric vascular bed is incubated with an enzyme solution containing collagenase, deoxyribonuclease, papain, dithiothreitol and bovine serum albumin for 45 min at 37 °C in a shaking waterbath. After the 45 min digestion, cells are centrifuged and plated. This method yields an endothelial cell population with a high plating efficiency which is relatively free of smooth muscle contamination.