Figure 8

Functional Assessment and Cell Signature of Preserved hiPSC-RPE Cell Suspensions. (a) hiPSC-RPE cell preservations of 6 and 24 hours period at varied temperatures were plated and then cell proliferation was evaluated by MTS assay each day from 1 to 6 days and compared to control populations (n = 9). (b,c) VEGF/PEDF secretion quantified by ELISA assay from varied preservations of hiPSC-RPE cells after 13 days in recovery culture and compared to control (n = 6). (d) Detection of RPE genes in hiPSC-RPE cells preserved at 16 °C for various periods. RT-qPCR of human RPE65, TYROSINASE, and PEDF represent (ΔΔCt to the 0 hour control sample) (n = 3). Mean ± SEM are presented. The one-way ANOVA with Tukey’s post hoc pair-wise comparisons test was performed. (e) Epithelial morphology in recovery culture of hiPSC-RPE cells preserved at 16 °C for various periods. Cells were plated into a 24-well CELLstart-coated plate after preservation and cultured for 21 days. Morphology was assessed by phase contrast and immunofluorescence microscopy (red, ZO-1). Scale bars = 50 μm.