Figure 3.

Newborn CD14 + monocytes are intrinsically impaired to extravasate quiescent but not inflamed endothelia. Monocytes were extracted from TCs after 1.5 h extravasation at 37ºC/5%CO₂ under either autologous plasma on quiescent endothelium, plasma-free on quiescent endothelium or plasma-free on pre-activated endothelium conditions, as described in methods. Pre-activation of endothelium was done with a cocktail of TNF-α, IL-1β, IL-6 and PGE₂ for 1 h (37 °C/5%CO₂). Extravasated monocytes were analysed by multicolour flow cytometry and the number of extravasated CD14 + monocytes (Cells/TC) was calculated by combining viable cell counts with the percentages of viable monocytes detected by flow cytometry (N = 3–6/age-group). Two-tailed Paired t-Test compares same study participant between conditions. Two-tailed UnPaired t-Test compares age groups. n.s. = non-significant; *P ≤ 0.05; ***P ≤ 0.005.