We present an approach for determining in vitro the means and distributions of a set of phenomenological parameters, including cell speed and persistence time, which can be used to evaluate the effect of isotropic variations in the extracellular environment on the motility of human tissue cells. Using time-lapse videomicroscopy and semi-automated image analysis, we tracked the paths traveled by slow-moving, isolated human vascular smooth muscle cells over 48 hours on surfaces of petri dishes coated with 10 μg/mL of the adhesive extracellular matrix proteins type IV collagen, fibronectin or laminin. By applying a persistent random walk model to experimental data for mean-squared displacement as a function of time for these cells, we rigorously distinguished individual cells with different motile characteristics not obvious based on qualitative comparisons between the structures of individual cell paths. We also positively identified the presence of immotile cells. Based on the behavior of 34 to 77 cells on each substrate, we found mean cell speeds and persistence times on the order of 10 micron/h and 3 hours, respectively, on all three ECM substrates, while the fraction of motile cells varied from 65% on laminin to 78% on collagen. On all three surfaces experimental number distributions of speed and persistence time could be described by normal and exponential waiting time distributions, respectively. Our approach provides a framework for addressing questions concerning the mechanistic relationship between cellular and environmental properties and cell motility.