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BMC Cardiovascular Disorders
Published in: BMC Cardiovascular Disorders 1/2007

Open Access 01-12-2007 | Technical advance

A cell culture model using rat coronary artery adventitial fibroblasts to measure collagen production

Authors: Cathleen Jenkins, Amy Milsted, Kathleen Doane, Gary Meszaros, Jonathan Toot, Daniel Ely

Published in: BMC Cardiovascular Disorders | Issue 1/2007

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Abstract

Background

We have developed a rat cell model for studying collagen type I production in coronary artery adventitial fibroblasts. Increased deposition of adventitial collagen type I leads to stiffening of the blood vessel, increased blood pressure, arteriosclerosis and coronary heart disease. Although the source and mechanism of collagen deposition is yet unknown, the adventitia appears to play a significant role. To demonstrate the application of our cell model, cultured adventitial fibroblasts were treated with sex hormones and the effect on collagen production measured.

Methods

Hearts (10–12 weeks) were harvested and the left anterior descending coronary artery (LAD) was isolated and removed. Tissue explants were cultured and cells (passages 2–4) were confirmed as fibroblasts using immunohistochemistry. Optimal conditions were determined for cell tissue harvest, timing, proliferation and culture conditions. Fibroblasts were exposed to 10-7 M testosterone or 10-7 M estrogen for 24 hours and either immunostained for collagen type I or subjected to ELISA.

Results

Results showed increased collagen staining in fibroblasts treated with testosterone compared to control and decreased staining with estrogen. ELISA results showed that testosterone increased collagen I by 20% whereas estrogen decreased collagen I by 15%.

Conclusion

Data demonstrates the usefulness of our cell model in studying the specific role of the adventitia apart from other blood vessel tissue in rat coronary arteries. Results suggest opposite effects of testosterone and estrogen on collagen synthesis in the rat coronary artery adventitial fibroblasts.
Appendix
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Metadata
Title
A cell culture model using rat coronary artery adventitial fibroblasts to measure collagen production
Authors
Cathleen Jenkins
Amy Milsted
Kathleen Doane
Gary Meszaros
Jonathan Toot
Daniel Ely
Publication date
01-12-2007
Publisher
BioMed Central
Published in
BMC Cardiovascular Disorders / Issue 1/2007
Electronic ISSN: 1471-2261
DOI
https://doi.org/10.1186/1471-2261-7-13

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