Tissue Engineering Using Vascular Organoids From Human Pluripotent Stem Cell Derived Mural Cell Phenotypes

Frontiers in Bioengineering and Biotechnology

Dimitrios Kouroupis¹  Eleni Bagli¹  Athena Kyrkou¹  Theodore Fotsis¹  Maria Markou¹  Carol Murphy¹  Athanasios Katsouras¹  Fotios Badounas³ 

[1] Foundation for Research and Technology-Hellas, Department of Biomedical Research, Institute of Molecular Biology and Biotechnology, Ioannina, Greece;Laboratory of Biological Chemistry, Medical School, University of Ioannina, Ioannina, Greece;Transgenic Technology Laboratory, Inflammation Group, Department of Immunology, Hellenic Pasteur Institute, Athens, Greece;

关键词: mural cells;    tissue engineering;    induced pluripotent stem cells;    smooth muscle cells;    vascularization;    vascular organoids;   

DOI  :  10.3389/fbioe.2020.00278

来源: DOAJ

【 摘 要 】

Diffusion is a limiting factor in regenerating large tissues (100–200 μm) due to reduced nutrient supply and waste removal leading to low viability of the regenerating cells as neovascularization of the implant by the host is a slow process. Thus, generating prevascularized tissue engineered constructs, in which endothelial (ECs) and mural (MCs) cells, such as smooth muscle cells (SMCs), and pericytes (PCs), are preassembled into functional in vitro vessels capable of rapidly connecting to the host vasculature could overcome this obstacle. Toward this purpose, using feeder-free and low serum conditions, we developed a simple, efficient and rapid in vitro approach to induce the differentiation of human pluripotent stem cells-hPSCs (human embryonic stem cells and human induced pluripotent stem cells) to defined SMC populations (contractile and synthetic hPSC-SMCs) by extensively characterizing the cellular phenotype (expression of CD44, CD73, CD105, NG2, PDGFRβ, and contractile proteins) and function of hPSC-SMCs. The latter were phenotypically and functionally stable for at least 8 passages, and could stabilize vessel formation and inhibit vessel network regression, when co-cultured with ECs in vitro. Subsequently, using a methylcellulose-based hydrogel system, we generated spheroids consisting of EC/hPSC-SMC (vascular organoids), which were extensively phenotypically characterized. Moreover, the vascular organoids served as focal starting points for the sprouting of capillary-like structures in vitro, whereas their delivery in vivo led to rapid generation of a complex functional vascular network. Finally, we investigated the vascularization potential of these vascular organoids, when embedded in hydrogels composed of defined extracellular components (collagen/fibrinogen/fibronectin) that can be used as scaffolds in tissue engineering applications. In summary, we developed a robust method for the generation of defined SMC phenotypes from hPSCs. Fabrication of vascularized tissue constructs using hPSC-SMC/EC vascular organoids embedded in chemically defined matrices is a significant step forward in tissue engineering and regenerative medicine.

【 授权许可】

Unknown