The transient receptor potential canonical (TRPC) 6 channel is an important ion channel located in podocytes, which plays an essential role in regulating calcium homeostasis of cell signaling. Podocytes are specialized, terminally differentiated cells surrounding glomerular capillaries, and are the subject of keen interest because of their key roles in kidney development and disease. Accordingly, we wonder whether TRPC6 channels undergo developmental changes during the podocyte differentiation and contribute to the maturation of podocytes. Using morphological, immunohistochemical and electrophysiological techniques, we investigated the changes of distribution and expression of TRPC6 in conditionally immortalized mouse podocyte cell line during differentiation. The results showed that the distribution of TRPC6 channels was changed with the maturity of podocyte differentiation, which was established by the morphological changes. TRPC6 proteins were mainly expressed in the nuclear regions of podocytes in undifferentiation state, and gradually translocated to other regions and increased in the cell membrane with the differentiation. Remarkably, the fluorescent intensity of TRPC6 cell surface significantly increased after the induction of differentiation. The data were corresponded with the increment of the density of current flowing through the channels recorded by whole-cell patch-clamp, indicating that the increased TRPC6 proteins on cell membrane were functional. The developmental changes of TRPC6 may contribute to the podocyte maturation and the physiological function. Our results have implications for the physiology and development of kidney and will serve as a baseline for future studies designed to investigate developmental changes of ion channel expression in podocytes.