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Updated: Mar 13, 2026
Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
Published on: November 20, 2017
Sex-related differences in matrix remodeling and early osteogenic markers in aortic valvular interstitial cells
Shirin Masjedi1, Ying Lei1, Jenny Patel1
1The Department of Mechanical, Aerospace and Biomedical Engineering, The University of Tennessee, 312 Perkins Hall, Knoxville, TN, 37996, USA.
Insights
Male aortic valve cells show greater calcification potential than female cells, indicating sex-based differences in calcific aortic valve disease (CAVD). This study explored differences in valvular interstitial cells (VICs) from male and female rats and pigs.
Area of Science:
- Cardiovascular Biology
- Cell Biology
- Biochemistry
Background:
- Calcific aortic valve disease (CAVD) is a significant cardiovascular condition.
- Male sex is a known risk factor for CAVD, but the underlying mechanisms remain unclear.
Purpose of the Study:
- To investigate sex-based differences in osteogenic differentiation and extracellular matrix (ECM) remodeling of valvular interstitial cells (VICs).
Main Methods:
- VICs were isolated from male and female rat and porcine aortic valves.
- Cells were cultured in osteogenic media for 10, 15, and 20 days.
- Proliferation, ECM remodeling (GAG, collagen I, MMP-2), and osteogenic markers (ALP, Alizarin Red S) were assessed.
Main Results:
- Female VICs exhibited greater proliferation than male VICs.
- Male VICs showed increased GAG, collagen I, and activated MMP-2.
- Male VICs displayed higher alkaline phosphatase (ALP) activity and formed larger calcific nodules.
Conclusions:
- Significant sex-related differences exist in aortic VIC osteogenic differentiation.
- Male VICs are more susceptible to calcification, contributing to CAVD pathogenesis.
Abstract:
Calcific aortic valve disease (CAVD) is a major cardiovascular disorder in the developed countries. Male is a known risk factor in this disease; unfortunately, how sex contributes to CAVD is mostly unknown. The objective of this study is to determine whether valvular interstitial cells (VICs) isolated from male versus female aortic valves demonstrate difference in osteogenic differentiation and/or extracellular matrix (ECM) remodeling. VICs were isolated from male and female rat or porcine aortic valves and cultured in osteogenic media for 10, 15 and 20 days. The proliferation among male and female VICs was assessed by a cell growth assay. The matrix remodeling of the VIC samples was quantified using glycosaminoglycan (GAG), collagen type I and gelatin zymography assays. Early osteogenic marker expression was assessed using alkaline phosphatase (ALP) staining and enzyme activity assay and Alizarin Red S staining. Our result showed that proliferation of VICs was significantly greater in female than male after 12 days of culture in regular media. Additionally, male VICs showed elevated amounts of normalized GAG, collagen I, and activated matrix metallopreoteniase-2 expression compared to female. Similarly, ALP content was greater in male VICs than female at all time points. In addition, male VICs formed calcific nodules with greater size, % area and integrated density than females. The results from this research suggest that there is a sex-related difference in the events associated with osteogenic differentiation of the aortic VICs, where male VICs are more prone to calcification.
