Dual In Vitro Perfusion of an Isolated Cotyledon as a Model to Study the Implication of Changes in the Third Trimester Placenta on Preeclampsia

Immunohistochemical detection of p22phox expression in control tissue at the beginning of the experiment (A, C) and after perfusion under standard (B), and prooxidant (X+XO) conditions (D). Arrows and arrowheads indicate trophoblast and Hofbauer cells respectively. Bar=100μm.

Immunohistochemical detection (i) and Western blot analysis (ii) of IL-1β expression in control tissue at the beginning of the experiment (A, C) and after perfusion under standard (B), and prooxidant (X+XO) conditions (D). Arrowheads indicate Hofbauer cells. Bar=100μm.

Total accumulation of 8-iso-PGF2α over 7h in the fetal (F), the maternal (M) and both circuits (M+F) under standard conditions (n=5) and in the presence of X+XO (n=4). Accumulation values calculated as mean±SD. *Standard vs. X+XO: P<0.04.

Flow cytometric analysis of the sediment of perfusion medium collected from the maternal circuit and stained for PLAP (i and iii) and from the fetal circuit and stained for CD62E (ii and iv). (i and ii): percentage of positive events in the three perfusate fractions; (iii and iv): representative analysis of the microparticles from the last perfusate fraction. Values are expressed as mean±SD of the percentage of the positive events.

(i) Immunohistochemical detection and (ii) Western blot analysis of catalase expression in control tissue at the beginning of the experiment (A, C) and after perfusion under standard (B), and prooxidant (X+XO) conditions (D). Bar=100μm. Arrows and arrowheads indicate trophoblast and Hofbauer cells respectively.

(i) Immunohistochemical detection and (ii) Western blot analysis of MnSOD expression in control tissue at the beginning of the experiment (A, C) and in placental tissue perfused for 7h under standard (B), and prooxidant (X+XO) conditions (D). Bar=100μm. Arrows indicate trophoblast cells and arrowheads indicate Hofbauer cells.

Western blot analysis of protein extracts of MPs collected from the maternal circuit at the end of perfusion under standard (Crtl) and pro-oxidant conditions (X+XO).