Arthrospira platensis cultivation produces a saline spent medium that must be treated to allow its reuse, thus saving water and avoiding environmental pollution. This study evaluates the association of flocculation followed by adsorption to treat the spent medium by applying different concentrations of granular activated carbon (GAC) and ferric chloride (F), and using different residence times (T). The simultaneous optimization of the independent variables GAC, F, and T was performed using both a 23 central composite design and a response surface methodology. The cells cultivated in the medium obtained after the optimal conditions of treatment (GAC = 54.2 g L−1, F = 10.0 mg L−1, and T = 30.8 min) provided the highest maximum cell concentration, Xm = 3140 ± 77 mg L−1 in 0.5 L Erlenmeyer flasks with the highest protein biomass content (44.9%). The treated medium in such conditions was also used in a 3.5 L tubular photobioreactor (PBR), reaching Xm = 4033 ± 110 mg L−1 and biomass with high contents of both protein (47.3 ± 2.6%) and chlorophyll (9.7 ± 0.3 mg g dry cell−1). Therefore, this study can contribute to diminishing costs of A. platensis production by reusing its culture medium and improving its biomass quality in PBRs.