With the advancement of molecular biology techniques, cell cultures have become an important platform for the production of recombinant proteins for therapeutic purposes, such as biopharmaceuticals. Recombinant erythropoietin (EPOhr) obtained by biotechnological processes is a biopharmaceutical used mainly in the treatment of anemia associated with chronic renal failure. EPOhr is produced in animal cell culture systems, primarily using the Chinese Hamster Ovary (CHO) cell line. Supplementation strategies of the culture medium with key components for cell metabolism are associated with the increase of specific growth rates and product synthesis, in addition to guaranteeing necessary attributes for the quality of the product. The supplements studied were evaluated from experimental planning providing tools that favor the understanding of the experimental data.