Additional file 1 of Novel 3D embryo implantation model within macroporous alginate scaffolds

Additional file 1: Supplementary data 1. Presto blue (PB) quantitative analysis of RL95–2 cell constructs. RL95–2 cell-seeded scaffolds were incubated for 2 h with 10% (v/v) PB reagent (in DMEM medium, supplemented with 10% (v/v) FCS). Then samples of 100 μL of the medium were transferred to a black bottom 96-well plate and fluorescent readings were obtained at excitation and emission wavelengths of 560 nm and 590 nm, respectively. A calibration curve was prepared to quantify viable cells. Cell numbers at each time point were calculated by using a calibration curve and were normalized to the number of cells seeded into the scaffolds. Supplementary Fig. 1. (A) RL95–2 cell viability after 1- and 2-weeks culture, by PB analysis. PB analysis of scaffolds, seeded with 0.5×106 cells showed no significant decrease throughout 2 weeks culture. (B) Analysis of scaffolds, seeded with 6.25 × 104 cells, one order of magnitude less than A, showed a steady cell number after 1 week of culture and a significant increase after 2 weeks of culture.