Recycling microalgae growth media is essential to reduce the water and nutrient demands, and increase the viability of biomass production. However, it is not clear whether the harvesting method affects the outcomes of medium recycling on culture performance. In this study, Limnospira platensis was grown in a semi-continuous mode with daily harvest and dilution (0.25 d−1) for 50 days with medium refresh (control), or recycle after 0.22 µm or 10 µm filtration, or centrifugation. The concentration of extracellular polymeric substances (EPS) in the medium was 4- to 6-fold higher during semi-continuous growth when compared to batch mode and the highest after centrifugation. Medium recycling reduced growth rates by 30 %, but productivity remained unaltered during the experiments (0.24 ± 0.02 g L−1 d−1). EPS extracted from recycled media had similar compositions (mainly proteins and hydrocarbons) and no influence in the viscosity of solutions. The biochemical composition and rheology of cell concentrates varied widely, especially after centrifugation. The harvesting method affected the partitioning between free EPS in the medium and EPS adhered to the cell wall. If carefully applied to the microalgae industry, these findings can lead to more circular and sustainable biomass production.