Smooth muscle cells (SMCs) play a crucial role in cardiovascular diseases. Proteomic analysis using two-dimensional gel electrophoresis (2DE) associated with mass spectrometry allows characterization of the proteome and secretome of human smooth muscle. The presence of a distinct SMC population in the arterial wall implies that under normal conditions, SMCs are phenotypically heterogeneous. Intracellular and secreted proteins from a primary culture of SMCs obtained from patients undergoing coronary bypass surgery were analyzed using 2DE in order to determine their specific features. The 2D reference maps show that SMCs are involved in a wide range of biological functions. They could constitute a useful tool for a wide range of investigators involved in vascular biology, allowing them to investigate SMC protein changes associated with cardiovascular disorders or environmental stimuli.