Figure 6.

Evaluation of medium, cell seeding density, and cell distribution for long-term bioreactor culture. (a) Growth curves for CAF-hTERT, 231 cells, and HMMEC-hTERT when grown in EC medium (top panel), low-serum EC medium (middle panel), or HMMEC medium (bottom panel) in 2D culture. Data are the mean of three replicate wells for each time point. (b) Cell density and (c) cell aggregation in perfused surrogates at 21 days when initial cell seeding concentration was 2.1 × 10⁶ cells/100 µL ECM (high) or 1.05 × 10⁶ cells/100 µL ECM (low) and surrogates were cultured in low-serum EC (0.05% FBS) or HMMEC media (5% FBS) (n = 4–6 surrogates per condition, unpaired t-test, *p ⩽ 0.05 and **p ⩽ 0.01). (d) Evaluation of the effect of rotation during ECM polymerization on cell distribution immediately following ECM polymerization (n = 5 surrogates per condition, unpaired t-test, *p ⩽ 0.05). (e) Cell distribution in perfused surrogates following perfusion for 21 days with HMMEC medium. The effect of rotation and cell seeding density (high versus low) were compared (n = 4 surrogates per condition, unpaired t-test). Data in b–e are the mean and SEM.