Fig. 5
Purification of AcLDL positive cells by cytofluorimetry allows the generation of highly pure RPE monolayers. a Representative image of cell sorting based on Dil-AcLDL labeling. RPE samples were incubated overnight with 10 μg/ml Dil-AcLDL. Cells were washed in DPBS, dissociated by Accutase treatment and filtered through a 40-μm cell strainer prior to cell sorting. Cells were sorted based on staining by Dil-AcLDL (red). Unlabeled cells from RPE samples were used as negative control (N.C.). b AcLDL-negative (left panel) and AcLDL-positive (right panel) cells were collected after sorting and seeded at a concentration of 105 cells/cm2 in transwell filters coated with growth factor reduced Matrigel. Cell morphology and persistence of AcLDL molecules in AcLDL-positive cells were assessed by brightfield and fluorescent microscopy, respectively, at days 1, 20, and 45. c TEER values were measured at days 1, 20, and 45 using an EVOM2 voltohmmeter. Data are expressed as mean ± SEM. *P < 0.05 [Student t test]. n = 3