Generation of Vascularized Neural Organoids by Co-culturing with Mesodermal Progenitor Cells

Abstract

SUMMARY Organoids are three-dimensional (3D) constructs generated in stem cell cultures and are thought to mimic tissue and organ development in situ . However, until recently, they often exclusively recapitulated the development of the organ`s parenchyma without the major components of the organ stroma. Here, we describe a protocol to incorporate stromal components, first of all blood vessels, by co-culturing with induced pluripotent stem cell-derived mesodermal progenitor cells. For complete details on the use and execution of this protocol, please refer to Wörsdörfer et al. (2019) .

Human iPSC-derived mesodermal progenitor cells preserve their vasculogenesis potential after extrusion and form hierarchically organized blood vessels

“…Once initiated, mesodermal cells give rise to generation of all vascular wall cell types such ECs, SMCs and MSCs/fibroblasts that in turn can interact with each other and orchestrate the further steps of vascular morphogenesis including a hierarchic vessel system containing both micro and macrovessels with multilayered vessel wall as demonstrated in this work. We recently used hiMPCs in organoid models where they also provided vascularization [16,51] underlining the basic vasculogenic potential of hiMPCs. Remarkably, in comparison to the organoids where we observed vessels with only one layer of peri-endothelial cells we achieved in the current study the formation of vessels with multilayered tunica media as demonstrated by histology and electron microscopic analyses.…”

Applications of Engineering Techniques in Microvasculature Design

“…Organoid vascularization in co-culture relies on culturing stem cells with other cell types such as HUVECs, mesodermal progenitor cells ( 45 ), and mesenchymal stem cells (MSCs) ( 46 ). MSC-based self-condensation allows for MSCs and HUVECs to form aggregates that develop into vascularized organoids.…”

“…Capillary networks as well require support from mural cells known as pericytes to assist in contractility and maturation. More recently, it has been demonstrated that mesodermal progenitor cells (MPCs) have the potential to form hierarchical vascular networks by differentiating into multiple cell types such as ECs, smooth muscle cells, and mural cells ( 45 , 49 ). Using MPCs for vascularized organoid and vascular network development can benefit from higher plasticity compared to the use of MSCs and HUVECs.…”

Vascularized organoids on a chip: strategies for engineering organoids with functional vasculature

“…However, despite the extensive vascular network in the mesodermal part, there were few vessel sprouts in the neural part. 45,46 Advances in gene engineering offer possible solutions to vascularize organoids via co-differentiation that circumvent the tedious work of finding appropriate cocktails of growth factors. After Morita et al reported their method for converting human fibroblast into functional endothelial cells via ETS transcription factor 2 (ETV2) in 2015, 47 the role of ETV2 in specifying cells to endothelial and hematopoietic lineages has been extensively studied and utilized.…”