STEMdiff Blood Vessel Organoid Kit

The STEMdiff™ Blood Vessel Organoid Kit from STEMCELL Technologies is designed for the differentiation of human pluripotent stem cells (hPSCs) to blood vessel organoids. Based on the formulation by Wimmer and Penninger and using technology licensed from the Institute of Molecular Biotechnology (IMBA), the kit is optimized to generate organoids that develop extensive vascular networks and strong endothelial cell-pericyte interactions, supporting more physiologically relevant 3D models. It delivers a higher yield of organoids compared to homemade protocols, with the option to scale up for high-throughput screening in a 96-well format, while the simple protocol eliminates the need for serum screening and labor-intensive protocol optimization.

In the workflow, hPSCs maintained in mTeSR™1 or mTeSR™ Plus are seeded as single cells in 6-well ultra-low attachment plates using the kit's aggregation medium. mTeSR™1-maintained hPSCs are seeded at 0.2 - 0.4 × 10⁶ cells/well and mTeSR™ Plus-maintained hPSCs at 0.1 - 0.2 × 10⁶ cells/well. After 1 - 2 days, differentiation is initiated by switching to mesoderm medium for 3 days. On day 4 - 5, vascular induction is performed with vascular induction medium for 2 days. The resulting aggregates are embedded in a collagen-Matrigel® sandwich, where they sprout and mature into vascular networks in maturation medium over 5 days, with free-floating organoids collectable at day 15 for downstream assays.

Generated organoids contain endothelial cells and pericytes. In mTeSR™1-maintained cell lines, organoids contain 42.92 ± 3.97% CD31+ cells and 43.10 ± 3.31% CD140b+ cells (n = 3 - 4), while in mTeSR™ Plus-maintained lines they contain 54.29 ± 3.72% CD31+ cells (n = 3). According to a third-party listing, the organoids include CD31+/CD34+/CD144+/KDR+ endothelial cells and PDGFR-β+/CD146+/SMA+/NG-2+ pericytes. The product is sold for research use only.