Figure 11

Secreted CC16 protein was measured by ELISA. CC16 secretion of three different hESC lines was compared. Secretion could be detected starting from two to four days after the induction of the ALI conditions. A similar secretion pattern over time could be measured in all three lines, with a peak in secretion level that is measured around day 8 followed by a decrease and stabilization further over time. (Figure 11A) CC16 secretion in samples from human embryonic stem cells (hESC) VUB07 cultured on porous membranes with the ALI differentiation protocol were compared to the NO ALI control samples and the "undifferentiated hESC control" (Figure 11B). All samples were measured in triplicate. No protein secretion could be detected in "No ALI control samples" till 20 days of culture. In the "spontaneous differentiation control" no secretion could be measured at any time. The minimum detectable concentration was <50 pg/mL.