FIG. 7.

Breast cancer cells (BCC) proliferation. BCC were cultured for 48 h with human umbilical vein endothelial cells in three-dimensional collagen or collagen + chondroitin sulfate (CS) gels for 48 h under static or shear conditions. (a) Total cell count for all generations. Cell generations are noted as (b) G1, (c) G2, and (d) G3 and higher. (e) Ki-67 proliferation marker immunocytochemistry confocal microscopy images of BCC. BCC spheroids embedded in 2.2 mg/ml collagen-only stiffness control cultured with human umbilical vein endothelial cells (HUVEC) on top of the gels and exposed to (Left) static conditions and (Middle) shear stress conditions. (Right) BCC spheroids embedded in 1.5 mg/ml collagen + 20 mg/ml CS cultured with HUVEC on top of the gels and exposed to static conditions. BCC were stained with CellTrace Far Red and Ki-67 proliferation marker (green). Bar Scale = 50 μm. Error bars show mean ± SEM, n = 2 biological samples. All static or all fluidic conditions were analyzed with a one-way ANOVA with Tukey's post-test. Tukey groups are shown for statistically significant results (p < 0.05). Identical extracellular matrix conditions exposed to static or shear conditions were analyzed with an unpaired Student's t-test. Bars connected with * represent statistical significance with an unpaired Student's t-test (p < 0.05). Scale bar = 50 μm.