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Directed self-assembly of spheroids into modular vascular beds for engineering large tissue constructs
Daniel T O Carvalho1,2, Tália Feijão1,2, Mariana I Neves1,2,3
1i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, Porto 4200-135, Portugal.
Biofabrication
|November 4, 2020
Summary
This study presents a simple method for rapidly creating artificial vascular beds using spheroids. The optimal ratio of mesenchymal stem/stromal cells (MSCs) to outgrowth endothelial cells (OECs) was found to be 1:1 for vascular bed development.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- Artificial vascular beds are crucial for tissue engineering, but current methods are complex and time-consuming.
- Optimizing spheroid properties before assembly is often overlooked in biofabrication.
Purpose of the Study:
- To develop a simple and rapid biomanufacturing strategy for spheroid-based vascular beds.
- To investigate the optimal ratio of mesenchymal stem/stromal cells (MSCs) to outgrowth endothelial cells (OECs) for spheroid formation and vascular bed development.
Main Methods:
- Spheroids were formed using MSCs and OECs at various ratios (10:1, 5:1, 1:1, 1:5) in non-adhesive microwells and cultured for 7 days.
- Spheroid properties, including cell organization, extracellular matrix (ECM) components, and endothelial cell (OEC) state, were monitored over time.
- A rapid, one-step method using non-adhesive moulds was employed for spheroid fusion into patch-like vascular beds.
Main Results:
- The optimal MSC/OEC ratio for OEC retention and organization within spheroids was 1:1.
- Spheroids underwent dynamic remodeling, with OECs forming peripheral monolayers stabilized by ECM and pericyte-like cells, leading to increased surface stiffness.
- OECs transitioned to a quiescent state, and their sprouting potential decreased over culture time, suggesting immature spheroids are more therapeutically relevant.
- Rapid formation and fusion of spheroids into uniform, square-shaped patches were achieved, creating vascular beds with high surface area and endothelial sprouting potential.
Conclusions:
- A simple and rapid biomanufacturing approach for spheroid-based vascular beds has been established.
- The 1:1 MSC/OEC ratio is optimal for spheroid development, leading to organized vascular structures.
- The developed vascular patches are scalable and possess significant endothelial sprouting potential, making them promising for modular tissue engineering.
