Electrospinning is perhaps the most flexible method to produce nanofibers nowadays. Polymeric solutions are capable of being converted into nanofibers with a wide variety of biomedical applications, from wound healing textiles to smart and complex drug delivery systems. The production of these biomedical nanofibers depends on the correct application of the working parameters. These parameters are divided in three main streams: i) the electrospinning equipment conditions, ii) the polymeric solution properties and iii) the ambient conditions. In this work, a detailed overview of the electrospinning working parameters and how those factors influence the final production of biomedical nanofibers is presented.