Figure 1

Flow cytometric analysis of peritubular endothelial cells isolated by MicroBeads. (A) Flow cytometry analysis of CD146-PE positive endothelial cells before (left panel), after MicroBeads isolation (middle panel), and after MicroBeads isolation with Fc-blocking and trypsin digestion (right panel). Shaded areas are respective isotype controls. (B) Flow cytometry analysis of blood cell (CD45) contamination when CD146-PE positive endothelial cells were isolated with MicroBeads (1:20) with (right panel) or without kidney perfusion (left panel). Shaded areas are respective isotype controls. (C) After perfusion, CD146-PE positive cells before (top left panel) and after MicroBeads isolation at different MicroBeads concentration: 1:5 (top middle panel), 1:20 (top right panel), 1:30 (bottom left panel) and 1:50 (bottom right panel). Shaded areas are respective isotype controls. (G) Statistical analysis of each individual experiment (N = 3). After modification of kidney perfusion and optimization of MicroBeads concentration, the purity of CD146 positive cells increased significantly from 7.5 ± 0.3 (before separation) to as high as 83.3 ± 1.7. Data are expressed as mean ± SEM with N = 3 for each experimental group. *P < 0.05, **P < 0.01, ***P < 0.001.