Alginate Encapsulation of Ovarian Follicles

For encapsulation of the OFs, 0.5% of sodium alginate (Sigma-Aldrich, St. Louis, MI, USA) solution was made of sodium alginate, and 2.0% of calcium chloride (Sigma-Aldrich, St. Louis, MI, USA) solution was made of calcium chloride. These solutions were disinfected at 120 °C for 30 min [16 (link)]. A single OF was transferred to individual drop of alginate with pipette, and then this alginate bead was placed in the encapsulation solution of calcium chloride for 2 min (Figure 1c). After encapsulation, bead was washed with MEM-α (1X) medium for CaCl₂ removal [14 (link),17 (link)]. 20 OFs were encapsulated and half of them were cultured in dishes and remaining half were cultured on chips.