Free-floating Long-term Vascularized Mesenchymal Organoids [RNA-seq]

The vasculature is essential for tissue function and pathology. Spheroid co-cultures of endothelial and mesenchymal stem/stromal cells (MSCs) form consistent structures, but the vascular components are short-lived. iPSC-derived vascular organoids can establish complex vasculature but often have cell heterogeneous variable maturation and low reproducibility. We presents consistently-formed, free-floating, long-term Vascularized Mesenchymal Organoids (VMOs), formed by co-culturing human umbilical vein endothelial cells (HUVECs) and MSCs in a pre-gelled minimal Matrigel scaffold. We studied organoid dynamics over 60 days culture time and used spheroids cultured for 14 days as control. Overall design: RNA was extracted from pooled samples (organoids) cultured for 3, 14, 28 and 60 days respectively. RNA was also extracted from day 14 spheroids (HUVEC + MSC no ECM scaffold)