Isolation and Culture of Pulmonary Vascular Smooth Muscle and Endothelial Cells

Experimental quality is directly proportional to cell quality. This might not be a valid mathematical equation, but it very succinctly and elegantly describes the work of biologists from all fields of research: molecular biology or cell biology, proliferation or apoptosis assays, patch clamp electrophysiology or flow cytometry. One element lies at the heart of the success of each of these procedures: viable and healthy cells. Many days and weeks of valuable experimentation time have been spent perfecting cell isolation techniques, simply to guarantee that we can gather reliable data. Although it is possible to purchase cells from various sources, most research groups have developed their own techniques for isolating cells from tissues, techniques which can be adapted relatively easily from one tissue type to another. In addition, although it is always desirable to use freshly isolated cells (less than 8 h after isolation), many investigators have turned to cell culture as a viable ­alternative to freshly dissociated cells, although this may present some scientific challenges and dilemmas. The current ­chapter addresses the isolation of pulmonary artery smooth ­muscle cells (PASMCs) and pulmonary artery endothelial (PAECs), particularly from humans, rats, and mice. Generally speaking, the methods we outline can also be applied to PASMCs and PAECs from other species, although some modifications may be required. Readers are advised to consult the extensive literature to identify the technique most applicable to their needs.