FIG. 3.

TEER of D8 cryopreserved cells. (A) Maximum TEER values of IMR90-4 iPSC-BMECs cryopreserved at D8 of differentiation. TEER values from five independent differentiations are shown, with frozen and nonfrozen samples paired from the same independent differentiation. Maximum values were recorded at 48 h post-thaw for each sample. Data for differentiations one to five represent the average ± standard deviation of triplicate Transwell filters from a single independent differentiation. The average ± standard deviation of the five independent differentiations is also depicted. Statistical significance was calculated through Student's unpaired t-test (**p < 0.01, ***p < 0.001, ****p < 0.0001). (B) Maximum TEER of IMR90-4 iPSC-BMECs as a function of time in cryostorage. IMR90-4-derived cells were cryopreserved at D8 of differentiation and stored in liquid nitrogen for various lengths of time. Each data point represents the average maximum TEER value from triplicate filters from a single independent differentiation. Symbols indicate different independent differentiations. Statistical significance was calculated through ANOVA (*p < 0.05). (C) (i) TEER profile of IMR90-4 iPSC-BMECs cryopreserved at D8 of differentiation with or without 10 μM Y-27632 treatment. Data represent average ± standard deviation from triplicate Transwell filters from one independent differentiation, and the experiment was repeated for two additional differentiations to verify trends. (ii) TEER from days 2, 4, and 6 post-thaw. Data are derived from (i) and replotted here to depict the relevant statistical comparisons. Statistical significance was calculated through ANOVA (**p < 0.01, ***p < 0.001, ****p < 0.0001). TEER, transendothelial electrical resistance.