A method for isolating and culturing vascular smooth muscle cells from porcine cerebral arteries

Cerebral blood vessels have unique properties when compared to most peripheral vascular beds. One such property is that large cerebral vessels are involved in the regulation of cerebral vascular resistance. Studying smooth muscle cells isolated from these vessels will determine how phenotypic properties of these cells contribute to unique cerebrovascular function. Therefore we developed a method of culturing smooth muscle cells from explants of cerebral arteries of porcine brains obtained, gratis, from a local slaughter house. Cells isolated and cultured by the methods described herein were of smooth muscle origin as indicated by histochemical staining for smooth muscle α-actin. Further, we examined the response of the cultured cells to agonists which activate the cGMP dependent vaso-dilator system by stimulating soluble guanylyl cyclase (nitroglycerin and sodium nitroprusside) or particulate guanylyl cyclase (C-type natriuretic peptide). Also, forskolin activation of adenylyl cyclase was examined. These agents stimulated an increase in intracellular cGMP and cAMP in a manner that was reproducible in every cell isolation (20 brains) and which remained unchanged through nine passages. Additionally, the cells could be frozen, thawed and replated without loss of responsiveness to these agents. The protocol reported here provides a method for culturing cerebral artery smooth muscle cells that is inexpensive, relatively simple, and which yields cells that can be utilized through multiple passages.