Background and Aims

Vascular smooth muscle cells (VSMCs) are known largely for their role in vasomotor regulation and maintenance of vascular homeostasis. Depending on microenvironment, VSMCs orchestrate their local architecture, leading to VSMC phenotypic modulation. Endothelial cells (ECs) are closely related to VSMCs of vessel wall with constant interactive communication between both cell types.

Phenotype modulation is one of the early signs of pathogenesis of atherosclerosis. Classically, monolayer VSMC cell culture models were used for disease modeling. However, these models lack three-dimensional (3D) structure and physiological features of blood vessels. Hence, we propose a technique to develop 3D vascular cell culture model system (spheroids) that would be more suitable for disease modeling with greater physiological relevance. Hence, a robust and reproducible vascular spheroid is needed for better disease modeling.

Methods

VSMCs and ECs were seeded into low attachment 96-well plate and maintained at 37 °C with 5% CO₂. The co-cultures were allowed to aggregate and self-formed to 3D vascular structure. For cellular confocal imaging, the spheroids were first labelled with Hoechst 33342 and subsequently labelled with VSMC markers (ACTA2 and Vimentin). Images were captured using High Content Imaging System.

Results

We observed that VSMCs and ECs co-culture aggregate and form 3D viable spheroids within 5 days of incubation.

Conclusions

The VSMC spheroid system is viable option to form 3D vascular model. It could be considering as a powerful platform that would be suitable as a tool to study vascular disease and as a platform to screening therapies.