Affiliation:
1. Laboratory of Experimental Cardiology University Medical Center Utrecht Utrecht the Netherlands
2. Central Diagnostics Laboratory University Medical Center Utrecht Utrecht the Netherlands
3. Department of Cardiology University Medical Center Utrecht Utrecht the Netherlands
4. Center for Translational Immunology University Medical Center Utrecht Utrecht the Netherlands
5. Pediatric Immunology and Rheumatology, Wilhelmina Children’s Hospital University Medical Center Utrecht Utrecht the Netherlands
6. Department of Vascular Surgery University Medical Center Utrecht Utrecht the Netherlands
Abstract
Background
Plaque myofibroblasts are critical players in the initiation and advancement of atherosclerotic disease. They are involved in the production of extracellular matrix, the formation of the fibrous cap, and the underlying lipidic core via modulation processes in response to different environmental cues. Despite clear phenotypic differences between myofibroblast cells and healthy vascular smooth muscle cells, smooth muscle cells are still widely used as a cellular model in atherosclerotic research.
Methods and Results
Here, we present a conditioned outgrowth method to isolate and culture myofibroblast cells from plaques. We obtained these cells from 27 donors (24 carotid and 3 femoral endarterectomies). We show that they keep their proliferative capacity for 8 passages, are transcriptionally stable, retain donor‐specific gene expression programs, and express extracellular matrix proteins (
FN1
,
COL1A1
, and
DCN
) and smooth muscle cell markers (
ACTA2
,
MYH11
, and
CNN1
). Single‐cell transcriptomics reveals that the cells in culture closely resemble the plaque myofibroblasts. Chromatin immunoprecipitation sequencing shows the presence of histone H3 lysine 4 dimethylation at the
MYH11
promoter, pointing to their smooth muscle cell origin. Finally, we demonstrated that plaque myofibroblasts can be efficiently transduced (>97%) and are capable of taking up oxidized low‐density lipoprotein and undergoing calcification.
Conclusions
In conclusion, we present a method to isolate and culture cells that retain plaque myofibroblast phenotypical and functional capabilities, making them a suitable in vitro model for studying selected mechanisms of atherosclerosis.
Publisher
Ovid Technologies (Wolters Kluwer Health)
Subject
Cardiology and Cardiovascular Medicine