Culture and extraction of S. maxima

Marine Spirulina maxima that has been originally cultivated in Korea Institute of Ocean Science & Technology (Jeju, Korea) and cultivated in vertical rounded-200 L-photobioreactor containing SOT medium with continuous aeration^19,57,58. Cultures were grown at 25 °C under a 12:12-h light/dark photoperiod. Cells were harvested by centrifugation at 9000 rpm for 20 min (Thermo Fisher Scientific, Massachusetts, USA) and then lyophilized and stored at − 50 °C (Operon, Gimpo, South Korea)⁵⁷.

Dried S. maxima powder was dissolved in distilled water (DW) at 1% (1×) and 0.5% (0.5×) (w/v), sonicated (Fig. ​(Fig.1A)1A) and subjected to high temperature and pressure treatment (Fig. ​(Fig.1B)1B) to disrupt the cells. The sample was centrifuged (Labogene, Daejeon, South Korea) at 9000 rpm for 20 min (Fig. 1C) and filtered through a 1-μm Whatman No. 1 filter paper (Whatman, Maidstone, England) (Fig. 1D). To separate the micro residues, the supernatant was centrifuged (Beckman Coulter, Brea, USA) at 30,000 rpm for 20 min (Fig. 1E) and filtered through a 0.2-μm filter to remove bacteria, fungi, and mycoplasma (Fig. 1F). The solutions [1% and 0.5% spirulina animal cell culture solution (SACCS)] were stored at − 20 °C until use. Under this process, the yields of SACCS extracted from Spirulina maxima was obtained by approximately 10%.