Human dermal fibroblasts were obtained from various anatomical sites expressing different sets of genes. Distinction of their synthetic can be determined by the microenvironment and composition of receptors on the surface of the cells. Earlier, we have shown that the cytoskeleton of cultured cells rapidly reacts to any external modification, and characteristics of their spatial organization may reflect similarities and differences between cells if various origins under the same conditions. The extracellular matrix proteins influence on the character of structures forming by actin cytoskeleton. In this work, we analyzed cytoskeleton organization of the fibroblasts obtained from normal postnatal, scar an embryonic human skin, which were spreading on the basic proteins of extracellular matrix such as collagen Type I and IV, laminin 2/4, and fibronectin. The results of confocal microscopy showed that fibroblasts of various origins had considerable differences in organization of actin structures and distribution of focal contacts visualized by anti-vinculin antibodies. Furthermore, various fibroblasts spread on the same protein of the extracellular matrix revealed essential modifications of actin structures and focal contacts with the similarity of spatial organization of the cytoskeleton. Discovered variations of actin microfilaments organization are evidence of different extent of cell interactions with various proteins. These variations may depend on combination of integrins on the surface of a cell membrane. The data obtained give grounds to suppose considerable differences in morphogenic function of fibroblasts of various origins.