Effect of ICG-001 on differentiation of hES cells in SAGM to AEI cells.

Single cell suspensions of H7 hES cells differentiated in SAGM were incubated with 5 µM ICG-001 in culture medium for 12 h and assayed by FACS to identify percentage of AEII cells (SP-C⁺), AEI cells (AQP-5⁺), and Clara cells (CC-10⁺). Percentage of total cells in culture of cells positive for a intracellular and b surface expression of AEI cell, AEII cell, and Clara cell markers. Surface expression of d pluripotent markers for hematopoietic cells (c-kit⁺), lineage-negative (Lin⁻) cells, and Oct3/4⁺ pluripotent cells, and e growth factors (EGF⁺, VEGF⁺, IGFII⁺). The percentage of positive cells of total cells in culture is shown as mean ± SEM (n = three independent experiments with FACS analyses performed in triplicate). P<0.05 values in ICG-001-treated group (open bars) vs. ICG-001-untreated group (solid bars) are shown. Representative FACS scattergrams with the percentage of cells gated in each quadrant shown for expression of c AQ-5 (UL, 0.03%; UR, 5.08%; LL, 4.92%; and LR, 89.97%) and e: Oct3/4 and VEGF (UL, 1.47%; UR, 0.53%; LL, 86.15%; and LR, 11.85%).