Figure 4.

Reduced HC and EC development from SDS-iPSC derived KDR⁺ CD34⁺ hemoangiogenic progenitors. (a) Outline of the differentiation protocol for generation of HCs and ECs from iPSC-derived hemoangiogenic progenitors. On day 6 of initial differentiation, sorted KDR⁺CD34⁺ cells were transferred onto fresh confluent OP9 cells, and were then cultured in the indicated culture conditions. (b) Isolation of KDR⁺CD34⁺ hemoangiogenic progenitors by FACS. Purities of viable KDR⁺CD34⁺ cells were 7.1% ± 1.3% in at least three independent experiments. (c) May–Giemsa staining of floating HCs obtained from SDS and control iPSC–derived KDR⁺CD34⁺ cells on day 26 of differentiation. Scale bar: 100 mm. (d, e) Sequential analysis of the number of floating HCs generated from SDS and control iPSCs (d), and from SDS-iPSC clones transduced with SBDS or empty vector (e). (f) Immunostaining of CD31 in EC clusters obtained from SDS and control iPSC–derived KDR⁺CD34⁺ cells on day 13 of differentiation. Scale bar: 500 μm. (g, h) Numbers (g) and area (h) of EC clusters generated from SDS-iPSC clones transduced with SBDS or empty vector, and from control iPSCs. Data represent means ± SEM of triplicate wells; representative results from one of three independent experiments are shown (*P < 0.05).