Generating Vascular Organoids

This is intended as a guide only; for full experimental details please read the reference provided.

Wimmer et al. describe a protocol to generate human blood vessel organoids from H9 ES cells or iPS cells.

Human H9 ES cells or iPS cells were disaggregated then resuspended in differentiation medium 1 containing Y-27632 (50 μM) and plated into one well of an ultra-low attachment six-well plate for cell aggregation. On day 3 cell aggregates were treated with 12 μM CHIR 99021, then on days 5, 7 and 9 BMP4, VEGF-A and FGF-2 were added.

On day 11, differentiation media 2 containing VEGF-A, FGF-2 and SB 431542 (10 μM) was added to increase the yield of endothelial cells and suppress pericyte formation.

On day 13 cell aggregates were embedded in Matrigel:collagen I (1:1) gels and differentiation media 3 was added. Media was changed every second to third day. Approximately day 18, vascular networks were established, extracted from the gels and further cultured in 96-well low-attachment plates. These vascular networks self-assembled into vascular organoids and could be cultured for up to 3 months.