The glomerular podocyte is a highly specialised cell, with the ability to ultrafiltrate blood, and support glomerular capillary pressures. However, little is known about either the genetic programs leading to this functionality, or the final phenotype. We approached this question utilising a human conditionally immortalised cell line, which differentiate from a proliferating epithelial phenotype to a differentiated form. We profiled gene expression during several time points during differentiation, and grouped the regulated genes into major functional categories. A novel category of genes that was upregulated during differentiation was of smooth muscle related molecules. We further examined the smooth muscle phenotype and showed that podocytes consistently express the differentiated smooth muscle markers smoothelin, calponin and the specific transcription factor myocardin, both in vitro and in vivo. The contractile contribution of the podocyte to the glomerular capillary is controversial. We demonstrated using two novel techniques that podocytes contract vigorously in vitro, when differentiated, and in real-time were able to demonstrate angiotensin II treatment decreases monolayer resistance, morphologically correlating with enhanced contractility. We conclude that the mature podocyte in vitro possesses functional apparatus of contractile smooth muscle cells, with potential implications for its in vivo ability to regulate glomerular dynamic and permeability characteristics.