Cell adhesion and migration behaviors are widely involved in physiological processes, and can be mediated by various factors. Collagen is one representative constituents of extracellular matrix, possessing advantages including excellent biocompatibility, low immunogenicity, and ease to be engineered, which makes collagen be promising biomaterial for regulating cell behaviors. Furthermore, the biofunction of collagen is closely correlated with its configuration. Whereas, current studies on collagen-based biomaterials for regulating cell adhesion and migration are rare, let alone the exploration of the ralationship between the configurations and these two behaviors. (Human umbilical vein endothelial cells, HUVEC) plays significant roles in wounds healing and cancer progression. Therefore, we aimed at disclosing the relationships between the adhesion/migration capability and the collagen origin, structure, and molecular state. In summary, results clearly revealed that denaturation disfavors the adhesion/migration of HUVEC cells, while assembly promotes. The expression levels of F-actin and vinculin, together with the quantity of filopodia were positively correlated with the two processes. Finnaly, our study may provide directions for the further designing of collagen-based biomaterials in the treatment of these kinds of severe diseases.