In this paper, the biomineralization potential and cellular response of novel blend films of the anionic sulfated polysaccharides kappa-carrageenan (KCG) and fucoidan (FUC) derived from seaweeds with semi-crystalline polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-valerate (PHBV), respectively, were analyzed. The incorporation of KCG and FUC into PHB and PHBV, which has been studied here for the first time, led to an overall decrease in crystallinity, enhanced surface hydrophilicity, reduced brittleness and faster degradation of the polymer blend films. All PHB/KCG, PHBV/KCG and PHBV/FUC films exhibited a two-stage mass loss profiles with pH stabilization. PHBV/KCG film showed the highest biomineralization activity due the presence of sulfate groups on the surface of the films. NIH3T3 cells attached and proliferated well on all blend films on account of enhanced surface hydrophilicity and improved flexibility. PHBV/KCG led to a promoted cellular activity compared to PHBV/FUC, presumably due to phase separation and higher amount of biopolymer on the film surface that was a consequence of the immiscibility of the polymers in the blend films.