Fig. 2: VEGF Captures Monocytes Under Flow.
From: Endothelialization of arterial vascular grafts by circulating monocytes
Microfluidic device to capture of cells from blood: schematic and shear dependency. a Schematic of microfluidic device. PDMS device is sealed to iVEGF using a vacuum. An empty channel image is shown. b VEGF binding kinetics to CH surface as determined by ELISA, n = 3 independent biological replicates; error bars denote ± SD of the mean. c MTT proliferation of HUVECs on iVEGF with a range of initial soluble VEGF concentrations added per well; n = 3 independent biological replicates; Four asterisk indicates statistical difference (p < 0.0001) compared to control (no VEGF) using one-way ANOVA and Dunnett’s test (DF = 12, F = 2101); error bars denote ±SD of the mean. d–f Whole human blood was passed through the microfluidic device on iVEGF at the indicated shear stress. All runs were 1 mL of blood. Total captured cells (d) were analyzed as captured cells per total cell count of 1 mL of blood, n = 10 independent biological replicates per shear strength; four asterisk indicates statistical difference (p < 0.0001), p-values as indicated determined by one-way ANOVA and Tukey’s test (DF = 36; F = 63.78); error bars denote ±SD of the mean. e, f Immunostaining with antibodies against MC markers CD14, CD163, and CD31 and EC marker CD144. e Representative images at 1 dyn/cm2 are shown. Images were quantified (f) as shown over the 10 independent biological replicates per shear (1 dyn; white bars, 5 dyn; diagonal thatched bars, 10 dyn; gray bars, 15 dyn horizontal thatched bars). No significant difference (p values indicated) between shear strengths and identity using two-way ANOVA and Sidak’s test; error bars denote ±SD of the mean. EC markers are observed in <1% of total cells under all shears tested. g VEGF captures cells expressing the VEGF receptors with high specificity, n = 10 independent biological runs per cell type (NIH-3T3; mouse fibroblast line, HF-MSC human hair follicle-derived mesenchymal stem cells, HD-FB human dermal fibroblasts, HUVEC human umbilical vein endothelial cell, OCAEC ovine carotid artery endothelial cell, RAW mouse macrophage line, H-MC human peripheral blood monocytes). Significance determined using two-way ANOVA and Sidak’s test (DF = 36, F = 1.458), significance as indicated; four asterisk denotes statistical difference p < 0.0001 as compared to cells captured on CH surface; error bars denote ±SD of the mean; iVEGF white bars, Hep surface gray bars.
