Human vascular smooth muscle cell culture

Human vascular smooth muscle cells (VSMCs) in culture are an important tool in understanding how VSMCs function and contribute to vessel wall contraction as well as disease. In this chapter, we describe methodologies that enable the investigator to culture large numbers of proliferative VSMCs. These VSMCs are heterogeneous and vary in size, shape, and proliferative capacity depending on the disease state and location of the vessel of origin. Therefore, we also describe techniques to validate their identity as bone fide VSMCs. Briefly, the methods include information on how to dissect the blood vessel to remove the medial layer containing VSMCs, as well as methods on how to propagate these cells, by either allowing VSMCs to migrate from the explanted medial tissue or by enzymatically dispersing the cells from the tissue. Both methods are suitable for culturing VSMCs derived from most vessel types with modifications of the enzyme dispersal method suitable for the isolation of microvessel VSMCs. An important feature of VSMCs in culture is that they lose many of their in vivo contractile properties and so model disease-associated VSMCs in the vessel wall rather than a non-proliferative contractile cell. To overcome this limitation, we also describe alternate methods that enable the study of cultured VSMCs in their contractile state by allowing the VSMCs to remain within an intact vessel ring. Overall, these procedures enable the investigator to undertake a diverse array of experimental assays on cultured VSMCs.