Bacterial cellulose (BC) and gelatin are well-known biomaterials. The novel bacterial cellulose/gelatin composite scaffolds were prepared using aqueous gelatin solution and bacterial cellulose excreted by Acetobacter xylinum. The prepared bacterial cellulose/gelatin scaffolds were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, mechanical test, swelling, and thermal studies. The morphology of these bacterial cellulose/gelatin scaffolds indicated that the gelatin molecules could penetrate well between the individual nanofibers of the bacterial cellulose. With the incorporation of gelatin in the bacterial cellulose, the crystallinity index tended to decrease while the thermal stability was improved. After the incorporation of gelatin in the bacterial cellulose, Young’s modulus of the composite was increased from 3.7 GPa to 3.9 GPa, while the tensile strength and strain at break point were decreased from 170 MPa (7.5%) to 114 MPa (4%), respectively. The swelling behavior test indicated that the water uptake capacity of the composite was only half of the pure bacterial cellulose. Cell adhesion studies were carried out using 3T3 fibroblast cells. The cells incubated with BC/gelatin scaffolds for 48 h were capable of forming cell adhesion and proliferation. It showed much better biocompatibility than pure bacterial cellulose. So, the prepared BC/gelatin scaffolds are bioactive and may be suitable for cell adhesion/attachment, suggesting that these scaffolds can be used for wound dressing or tissue engineering scaffolds. Therefore, these novel BC/gelatin scaffolds are useful for biomedical applications.