This chapter demonstrates that electrospinning is a potential single-step method for producing three-dimensional (3D) scaffolds of tailored topological design mimicking the morphology of extracellular matrix (ECM). The electrospinning apparatus consists of a high-voltage power supply that can provide as much as 40 kV, with two electrodes, metering pump, a glass syringe with a small-diameter needle, and a collector. Specimens were prepared for scanning electron microscopy (SEM) examination by gold coating for 15 seconds. The structure of cellulose can be identified as a long-chain polysaccharide. Cellulose acetate (CA), one of the cellulose derivatives, can be dissolved in organic solvents such as acetone and acetic acid. Electrospinning can generate polymeric fibers with diameter ranging from a few nanometers to several tens of micrometers, and structures with open porosity. To produce the structure of Urinary Bladder Matrix (UBM) scaffolds, different CA solutions and processing parameters were designed, according to the knowledge obtained from the preliminary experiments with the single layer mats.