Engineering In Vitro Differentiation Microenvironments to Study Heart Valve Calcification

Yang, Y.; Bhunia, S.; Kontronias, R.; Qamar, S.; Srinivasan, L.; Ipek, T.; Deegan, A.; Lyu, L.

Abstract

Valvular calcification is one of the most common cardiac conditions affecting the elderly population. The reasons behind the calcification process and the cellular mechanisms involved are not well understood. Valve interstitial cells (VICs) are predominant cell type of cardiac valves with high heterogeneity in cell phenotype. We hypothesised that through engineering in vitro microenvironment, one VIC subtype can activate and differentiate into osteoblast-like cells. Thus, the activation process and the isolated subgroup cells can be developed into a facile technique to study the cause of heart valve calcification, the effect of aging on the calcification process and the effect of new drugs on calcification. In this study, primary aortic VICs have been isolated by sequential dissection of porcine aortic valves. The digested VICs were cultured on a specially treated substrate which effectively separated the VICs into distinct subgroups with different morphologies. The subgroup with cell cluster morphology stained strongly for Alizarin Red, von Kossa and E-cadherin. The subgroup prevalence increased with passage number under both osteogenic media and basal media, with a more considerable increase under osteogenic media. Clonal experiments confirmed the observation. PCA analysis via synchrotron-sourced microFTIR demonstrated that the cell cluster subgroup had a different spectral signature in comparison to the other subgroup; e.g. in monolayered cells, with increasing passage number expression difference in lipid and phosphate regions concomitantly increased. A preliminary drug assay using atorvastatin in the culture system revealed that the cell cluster's size was reduced and the shape assumed a less dense morphology.

Citation

Yang, Y., Bhunia, S., Kontronias, R., Qamar, S., Srinivasan, L., Ipek, T., …Lyu, L. (2015). Engineering In Vitro Differentiation Microenvironments to Study Heart Valve Calcification.

• Conference Name: 2015 4th TERMIS World Congress
• Conference Location: Boston, Massachusetts, USA
• Start Date: Sep 8, 2015
• End Date: Sep 11, 2015
• Online Publication Date: Aug 28, 2015
• Publication Date: Sep 1, 2015
• Deposit Date: Jun 9, 2023
• Publisher: Mary Ann Liebert
• Volume: 21 (S1)
• Publisher URL: https://www.liebertpub.com/doi/10.1089/ten.tea.2015.5000.abstracts