(A) Schematic of differentiation protocol used for vascular optimization.
(B) One well of a 96-well organoid plate treated with 100 ng/ml VEGF, showing podocytes (SYNPO), proximal tubules (LTL), and EC (CD31) by wide-field immunofluorescence.
(C) Wide-field images of VE-cadherin immunofluorescence in organoid cultures ± VEGF (left) or EC-directed cultures (right).
(D) Percentage of the total culture area occupied by cells expressing VE-cadherin, averaged from four representative experiments, or (E) expressing CD31, averaged from two additional representative experiments (± stderr).
(F) Confocal optical sections showing EC (CD31+) in optimized organoids, compared to human kidney sections.
Scale bars, 200 µm.