Abstract: Disclosed is a glycosaminoglycan-polycation complex as a matrix material for use in tissue regeneration, such as cartilage repair, and a preparation method thereof. The complex is synthesized by crosslinking glycosaminoglycan and polycation with a polyfunctional crosslinking agent under physiological conditions. In this method, the crosslinking agent consists of polyethyleneglycol having two or more electrophilic leaving groups (e.g. succinimidyl group or its derivatives) at the carboxyl terminal thereof, and the concentration of the crosslinking agent is in the range of 0.3 to 3 mM. According the present invention, the crosslinking reaction can be performed under physiological conditions to avoid conventional problems of adverse affect on cells (cell death) and the formation of a polyion complex (inhomogeneous precipitate) due to a crosslinking reaction in alcohol or water. Thus, the crosslinking reaction can be conducted in the presence of cells mixed to the glycosaminoglycan and polycation in advance.