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In order to explore the possibility of using bacterial cellulose (BC) as scaffold in tissue engineering, fibroblasts (FB) were cultured as seed cells on hydrated membrane of bacterial cellulose (BC). First, FBs were isolated by dispase-collagenase system from the dermis of the back skin of newborn (within 24 hours) rats and were adherently cultured in DMEM medium with newborn calf serum (10%) under 5% CO2 and 37°C at a density of 1.0×105 cell/ml. The growth curve of FBs was determined by MTT method. Second, the subcultured rat FBs were inoculated on hydrated membrane of BC as scaffold, at a density of 3.0×105 cell/ml. At different time intervals, the growth of cells on scaffold was determined by morphological characterization, such as HE staining, optical microscope, and scanning electron microscope (SEM). From the third to the seventh day of culture, the rat FBs were in logarithmic growth phase and showed strong proliferation, which might be well suited to be seed cells cultured on BC scaffold. After the inoculation for 3 days, rat FBs entered into the scaffold, and integrated closely with BC, which supported proliferation of cells. Excellent biocompatibility was verified in our experiment. The results suggested the potential for this biomaterial as a scaffold for tissue engineering.