Use of multiplied viable cells from the nucleus pulposus in altered discs, following in vitro cultivation, may be a promising therapy for degenerative disc disease. Up till now, alginate has been used as a three-dimensional cell carrier to cultivate nucleus pulposus cells. However, the biocompatibility of the alginate, which depends on the composition and purity of alginate materials used, has not been considered for in vivo application so far. In this study, we cultured porcine cells from the nucleus pulposus in a mixture of fibrin and hyaluronic acid (HA). The DNA content and proteoglycan synthesis were compared to those measured in an alginate matrix. Although the increase in DNA content was 2.5-fold higher in the alginate culture after 3 weeks, the proteoglycan synthesis in relation to the DNA content was significantly higher in the fibrin/HA matrix. We found that the fibrin/hyaluronic acid matrix can be used as a substrate for in vitro cultivation.