Investigation of differentially expressed genes in subpopulations of adherent human umbilical vein endothelial cells (HUVECS) isolated after fluid shear stress compared to the whole population of endothelial cells
The goal of this study is to determine whether EC loss is a random process or whether it is possible to predict which cells are more likely to remain adherent upon exposure to fluid SS. We used fluid SS to isolate a subpopulation of ECs that resist detachment and remain adherent. We used RNA-seq to examine differential gene expression of adherent subpopulation ECs compared to the whole population to identify targets for improving retention Overall design: Primary HUVECs were cultured overnight on fibronectin-coated 100mm dishes in the flow path area using a rectangular gasket and then subjected to 15 dyn/cm2 fluid SS using a rectangular parallel pate flow chamber. HUVECs were exposed to fluid SS for 0, 5, 10, or 30 min to generate the whole population, 5 min adherent, 10 min adherent, and 30 min adherent subpopulations, respectively. Plates from each time point were pooled together to generate sufficient RNA for analysis. A total of n=3 replicates from independent experiments were used for each time point.