This paper reports new experimental data of microalgae growth and lipid production under autotrophic conditions for the species Nannochloropsis salina. The effect of relevant operating variables is addressed and discussed, and some suggestions to better understand the process behavior are given with respect to lipid content maximization, carbon dioxide and nitrogen supply, and illumination conditions. The data obtained are finalized to the design of an environmentally and economically sustainable photobioreactor in view of achieving industrial photosynthetic biomass and natural oil production from large scale microalgae cultivation.