Fig. 5
BMEC differentiation using E6 medium translates to additional iPSC lines. iPSC lines CD12, CC3, and SM14 were differentiated to BMECs with E6 medium as described in Fig. 1, and TEER was measured approximately every 24 h. For each differentiation, three filters were seeded with BMECs, and each filter was measured at three locations on the filter. Each plot is the result of one biological replicate (n = 1) with each daily TEER measurement the result of a technical n = 9. All values are mean ± standard deviation of these nine total measurements per condition. a CD12-derived BMECs achieved maximum TEER values exceeding 4000 Ω × cm2 and maintained TEER above 1000 Ω × cm2 for a minimum of 11 days in 3 independent biological replicates. b CC3-derived BMECs achieved maximum TEER values exceeding 3500 Ω × cm2 and maintained TEER above 1000 Ω × cm2 for a minimum of 8 days in 3 independent biological replicates. c SM14-derived BMECs achieved maximum TEER values exceeding 2500 Ω × cm2 and maintained TEER above 1000 Ω × cm2 for a minimum of 11 days in 3 independent biological replicates. d Apical-to-basolateral flux of rhodamine 123 (R123) and H2DCFDA was measured across BMECs in the presence or absence of PSC833 and MK-571, respectively. Fluorescence was normalized to cells not treated with inhibitor and reported as normalized mean fluorescence ± standard deviation. Each condition was performed using triplicate filters and statistics were calculated using a technical n of 3. Statistical significance was determined using the Student’s unpaired t test (*p < 0.1; **p < 0.05; ***p < 0.01). Data from one biological experiment are shown, and an additional biological replicate was performed for each line to verify the observed trends. e The permeability of sodium fluorescein was measured across BMECs. Each experiment was performed using triplicate filters and data are presented as mean ± standard deviation. Biological duplicates were used to verify each measurement. The effective permeability (Pe) was calculated at less than 1.95 × 10−7 cm/s for all lines