Figure 3
From: Culture surface protein coatings affect the barrier properties and calcium signalling of hESC-RPE
POS phagocytosis assay performed on hESC-RPE 13 weeks post-seeding with purified porcine POS (hESC-08/017, Table 1). (a) Representative laser scanning confocal microscopy z-maximum intensity y projections (z-MIP) and xz cross-sections (MIP from 20 sections) after 2-h phagocytic challenge showing the overall number, distribution and internalisation of the POS particles. The POS particles were labelled with anti-opsin (green) and filamentous actin with phalloidin (red). Scale bar 20 μm. Quantification of internalised POS particles (b) per cell and (c) per field. Each field contains cs. 100 cells. Internalised POS particles were manually calculated from each xz-MIP image (from five z-MIP images/coating). Both data represent means ± SD. The number of POS particles per cell between LN + Col and LN + Col + 1xNid (p = 0.0079) and LN + Col + 10xNid (p = 0.0079) was statistically significant. Similarly, POS number per field on Col + LN was statistically significant compared to LN + Col + 1xNid (p = 0.0079) and LN + Col + 10xNid (p = 0.0159). Statistical analysis was performed with Mann–Whitney U test. (d) A schematic image to demonstrate which POS particles (green) were calculated as internalised POS particles (white arrows). Col—collagen IV, LN—laminin, Nid—nidogen-1.
