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Updated: Sep 1, 2025
Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation
Published on: May 31, 2016
Induced pluripotent stem cell-derived smooth muscle cells to study cardiovascular calcification
Samantha K Atkins1, Abhijeet R Sonawane1,2, Romi Brouwhuis1
1Center for Interdisciplinary Cardiovascular Sciences, Division of Cardiovascular Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, United States.
Insights
Researchers developed a new method using induced pluripotent stem cells (iPSCs) to create vascular smooth muscle cells (SMCs) for studying cardiovascular calcification, offering a scalable cell source for research.
Area of Science:
- Stem Cell Biology
- Cardiovascular Research
- Proteomics
Background:
- Cardiovascular calcification is a major cause of death and disease worldwide.
- Current treatments for cardiovascular calcification are limited to invasive surgical options.
- Vascular smooth muscle cells (SMCs) play a critical role in the development of vascular calcification.
Purpose of the Study:
- To establish an induced pluripotent stem cell (iPSC)-derived model of vascular smooth muscle cells (SMCs) for studying cardiovascular calcification.
- To characterize the proteomic changes during iPSC differentiation into SMCs (iSMCs).
- To develop an in vitro model for investigating vascular calcification using iSMCs.
Main Methods:
- Differentiated human iPSCs into iSMCs using a 10-day protocol.
- Characterized iSMC differentiation via morphology, immunofluorescence, flow cytometry, and proteomics.
- Induced calcification in iSMCs by culturing in osteogenic media for 17 days, verified by Alizarin Red S staining and proteomics.
Main Results:
- Successfully differentiated iPSCs into iSMCs, confirmed by multiple analytical methods.
- Proteomic analysis revealed key cellular pathway dynamics during iSMC development and showed high correlation with primary human SMCs.
- Established a functional in vitro model of vascular calcification using iSMCs.
Conclusions:
- Generated an inexhaustible source of functional vascular SMCs and calcifying SMCs from iPSCs.
- The iPSC-derived SMC model provides a valuable tool for studying cardiovascular calcification pathogenesis.
- This model holds significant potential for future cardiovascular calcification research and therapeutic development.
Abstract:
Cardiovascular calcification is the lead predictor of cardiovascular events and the top cause of morbidity and mortality worldwide. To date, only invasive surgical options are available to treat cardiovascular calcification despite the growing understanding of underlying pathological mechanisms. Key players in vascular calcification are vascular smooth muscle cells (SMCs), which transform into calcifying SMCs and secrete mineralizing extracellular vesicles that form microcalcifications, subsequently increasing plaque instability and consequential plaque rupture. There is an increasing, practical need for a large scale and inexhaustible source of functional SMCs. Here we describe an induced pluripotent stem cell (iPSC)-derived model of SMCs by differentiating iPSCs toward SMCs to study the pathogenesis of vascular calcification. Specifically, we characterize the proteome during iPSC differentiation to better understand the cellular dynamics during this process. First, we differentiated human iPSCs toward an induced-SMC (iSMC) phenotype in a 10-day protocol. The success of iSMC differentiation was demonstrated through morphological analysis, immunofluorescent staining, flow cytometry, and proteomics characterization. Proteomics was performed throughout the entire differentiation time course to provide a robust, well-defined starting and ending cell population. Proteomics data verified iPSC differentiation to iSMCs, and functional enrichment of proteins on different days showed the key pathways changing during iSMC development. Proteomics comparison with primary human SMCs showed a high correlation with iSMCs. After iSMC differentiation, we initiated calcification in the iSMCs by culturing the cells in osteogenic media for 17 days. Calcification was verified using Alizarin Red S staining and proteomics data analysis. This study presents an inexhaustible source of functional vascular SMCs and calcifying vascular SMCs to create an in vitro model of vascular calcification in osteogenic conditions, with high potential for future applications in cardiovascular calcification research.
