A public health problem is the loss of tissue due to different etiological factors. Then, a traditional way to rebuild lost tissue is by culturing cells onto the surface of biomaterials. These cultures have been carried out in extended surfaces which do not guarantee the cells growth. Nevertheless, in the last years, the culture over scaffolds arises as a technique which improves momentum, heat, and mass transfer. The biomaterial of these scaffolds is a choice which establishes the good or bad response from the host cells, and the procedure conditions to engineer these scaffolds depend on it. Thus, this manuscript aims to stablish the laboratory conditions to produce chitosan scaffolds. Promptly, it focuses on the study of the conditions to generate microcarriers by using an electrospray equipment, evaluating the physical and cytotoxic characteristics of the microcarriers. The polymer solution was discharged onto a polyphosphate solution for crosslinking the polymer. It was employed a factorial experimental design of three parameters: voltage, polymer concentration and solution flow rate. Microcarriers obtained were collected in a polyphosphate solution, and washed in a Hank´s solution. Microcarriers were analyzed in a light microscope in order to obtain the size distribution. Results evidence a hard relationship between voltage and the mean diameter of the particles, with a none influence from the flow rate nor polymer concentration. The best condition was with a voltage of 17 kV, polymer concentration of 0.5% w/v and solution flow rate of 4.5 mL/h. The mean particle diameter for these conditions was 568.04 ± 81.68 µm. Scanning electron microscopy (SEM) depicts low porosity, which allows the cell culture over the microcarriers surface. Finally, cytotoxicity of the microcarriers was evaluated by using a fibroblasts cell line without adverse effect on the cells. In this way, microcarriers may be used for culture of anchorage dependent cells in spinner bioreactors.