(a) Viability of MCF7 cells in MCF7-S, MCF7-EF-S, P-hTMES and I-hTMES, encapsulated in Alg or Alg/Alg-S following 96 hr incubation with constant media perfusion. (b) Imaging of P-hTMES, in Alg (upper panel) and Alg/Alg-S (lower panel), with MCF7 labeled with CFSE (green), fibroblasts with CMAC (blue), and dead cells with EtHD (red). Imaging was conducted using epifluorescence microscopy, using 20X maginification and 2 μm Z-stacking. Scale bar = 50 μm. (c) MCF7 cells viability following 24 hr perfusion with 5 μM Dox; or (d) 10 μM Dox. MCF7 cell viability is presented as mean viability ± S.D., n=3, calculated as the percentage of live cells in 50 scaffolds for each scaffold type, analyzed using ImageJ. Viability in the 2D culture was analyzed using Presto Blue, mean viability ± S.D., n=3. Statistical analysis in (a) and (c)-(d) was conducted by 2-way ANOVA, and one-way ANOVA, respectively, with p ≤ 0.05 considered statistically significant.