Overview of the sequential enzymatic digestion procedure for isolating cells from human aortic tissues.

<p>(<b>A</b>) The various steps of the isolation protocol are displayed. (<b>B</b>) A representative aneurysmal aorta segment is shown before and after longitudinal sectioning from its convex to its concave border. (<b>C</b>) A schematic diagram summarizes the fractionation procedure from the first round of enzymatic digestion (P0) to the expanded population (P2). (<b>D</b>) Histological analysis (H and E) of the representative aorta of patient 3, before and after collagenase digestion of the intimal or adventitial sides shows the efficiency of the enzyme action (4 rounds, 10 minutes each). (<b>E</b>) Phase contrast image of the crude cell population isolated after the first round of enzymatic digestion seeded in the 4-well plate (T0). Below left, a schematic representation of these cells seen from the top and from the side with the unattached cells floating in the culture medium.