The study of freshwater microalgae Audouinella pygmaea was performed using a bench bioreactor in different culture conditions to verify the best configuration in its growth and oil production, involving pH, dissolved oxygen, temperature control (20ºC and 28ºC), nutrient variation (nitrate and phosphate), and the insertion of CO2 (112 and 120 mg/L) in a reaction time of 96h. Two extraction methods, Soxhlet and Hara-Radin, were compared, and the last one was applied in the experiments.