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

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 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%.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.Cardiovascular disease is the number one cause of death in the United States. Diseases such as hypertension and coronary artery disease develop through multiple mechanisms; of significance are the structural changes that take place within blood vessels. In vivo data has shown that coronary artery fibrosis can result as a consequence of high blood pressure (BP). In addition to elevated BP, testosterone has been shown to increase the amount of collagen measured around coronary blood vessels contributing to fibrosis. For example, Seachrist found that testosterone increased coronary adventitial collagen in male hypertensive rats independent of the renin-angiotensin system [1]. Studies have provided further evidence for the role of testosterone in coronary artery fibrosis in male borderline hypertensive rats. Sympathectomy reduced systolic BP, but more interestingly reduced plasma testosterone levels correlated with decreased coronary artery collagen deposi