Presenter of 1 Presentation
UNDERSTANDING 3D CULTURE IN SMOOTH MUSCLE CELL BIOLOGY AND THE MODELLING OF ATHEROSCLEROSIS
Abstract
Background and Aims
Even though the advantages of 3D culture in mimicking cellular environment and drug discovery is widely reported in cancer biology, there are limited studies on 3D culture in vascular biology and atherosclerosis modelling. Our study aims to investigate the significance of 3D spheroid culture on cellular physiology and in atherosclerosis modelling using vascular smooth muscle cells(VSMC). We also explore 3D vessel modelling via co-culturing of endothelial cells(EC) and VSMCs.
Methods
Commercial SMC and patient-derived SMCs(CD45-/CD31-/CD90-), which were isolated using enzymatic dissociation and magnetic microbeads sorting from primary arterial tissue, were used.
Results
Transcript evaluation identified that SMCs in their adherent 2D culture and 3D floating culture (spheroid) have differentially expressed genes related to cellular phenotype and their response to oxLDL stimuli. 3D SMCs have significantly higher expression of genes encoding for foam cell-like characteristics(CD36 and LOX-1) and extracellular matrix-related proteins(MMP2, Fibronectin, and VCAM-1) compared to 2D VSMCs. Moreover, upon introducing ECs to VSMC 3D culture, we observed specific movement and self-organization with EC localization within the core region of spheroids, regardless of the time of EC introduction and cell ratio.
Conclusions
Thus, we conclude that transcripts differences of the 2D and 3D culture vascular could be utilized to create a patient-specific model, allowing precise therapy planning and drug testing ex vivo. The 3D vessel model could generate patient-specific disease model and provide greater understanding of VSMCs characteristic in the cellular microenvironment mimicking vessel wall and atherosclerosis. Moreover, the proposed 3D model would provide insights into the ECs and SMCs interaction on single-cell resolution.