Programmatic Introduction of Parenchymal Cell Types in Blood Vessel Organoids

We developed a bottom-up engineering approach to building vascularized human tissue by combining genetic reprogramming with chemically directed organoid differentiation. As a proof of concept, we created neuro-vascular and myo-vascular organoids via transcription factor overexpression in vascular organoids. We characterized neuro-vascular organoids (iN-VOs) by single cell RNA-sequencing (scRNA-seq) to determine cell type composition and gene expression. Using droplet based scRNA-seq we analyzed over 26,000 cells across day 45 organoids grown under three conditions: 45 days of NEUROD1 overexpression (iN-VO, 45 Days Induction), 15 days of NEUROD1 overexpression (iN-VO, 15 Days Induction) and without overexpression (iN-VO, No Induction). We find that organoids contain a neural cell population only upon induction of NEUROD1 while maintaining the full set of endothelial and mesenchymal lineages present in uninduced vascular organoids. Overall design: Single cell RNA-Seq for blood vessel organoids with 45 days, 15 days and no NEUROD1 induction