Development and characterization of a cloned rat pulmonary arterial smooth muscle cell line that maintains differentiated properties through multiple subcultures

Abstract

Background. Pulmonary hypertension is associated with abnormal pulmonary arterial contractility and growth. The mechanisms for these abnormalities are largely unknown. To study these processes, we sought to develop an in vitro system. Even though cultured aortic and pulmonary artery smooth muscle cells (SMCs) have been of considerable value in studying smooth muscle biology, one drawback of this system has been that these cells often lose differentiated properties in an unpredictable manner when they are passaged in culture. In addition, there appear to be significant differences in physiological and pathological responses between the systemic and pulmonary circulations, many of which could be directly related to the smooth muscle. We therefore established a cloned population of rat pulmonary arterial SMCs (PASMCs) that maintain differentiated properties through multiple subcultures. Methods and Results. PASMCs were obtained initially by enzymatic dissociation from pulmonary arteries of adult Sprague-Dawley rats. From these cells, clones were isolated. The cloned cells retained expression of functional surface receptors for angiotensin II, norepinephrine, and α-thrombin and high levels of the smooth muscle isoforms of α-actin, myosin heavy chain, myosin regulatory light chain, and α-tropomyosin mRNAs even after multiple passages. The cells could also be transfected and processed exogenous transcripts in a smooth muscle-specific fashion. Conclusions. These cloned PASMCs retain many differentiated characteristics and should be valuable for future studies of pulmonary vascular smooth muscle cell biology.