PMC

Cell population (mean ± standard deviation) over the course of six days while in three-dimensional culture (n = 12 per group). Culturing the cells within the differing three-dimensional environments did not produce any particular trend in the cell population. Statistical significance was not observed at the P = .05 level.

Extracellular pyruvate production after the final two days of two-dimensional culture (n = 12 per group). The production data does not show a clear trend. (a: significantly different from corresponding value in the 5% group; b: significantly different from corresponding value in the 10% group; e: significantly different from all other values within same serum group; f: significantly different from 5.0 mM value within same serum group; g: significantly different from 25.0 mM value within same serum group; h: significantly different from the 1.0 mM value within the same serum group; P = .05).

Fluorescence microscopy images indicating the viability of BMSCs in three-dimensional culture after the second and sixth days (increasing glucose concentrations from left to right; increasing collagen densities from top to bottom). Calcein-AM indicates living cells by emitting green light, while ethidium bromide indicates dead cells by emitting red light. After day 2, the 0.5 mM environment resulted in low viability, while the 25.0 mM environment resulted in high viability. By day 6, the 0.5 mM environment and the 25.0 mM−2.5 mg/mL group resulted in low viability, while the 25.0 mM−5.0 mg/mL group resulted in high viability and confluency. (40x objective; scale bar represents 100 μm).

Cell population (mean ± standard deviation) over the course of six days while in two-dimensional culture (n = 12 per group). Higher glucose and serum concentrations appear to produce higher cell populations over time. (a: significance between corresponding value in the 5% group; b: significance between corresponding value in the 10% group; c: significance between corresponding values in the 5% and 10% groups; d: significantly difference between the 5.0 and 25.0 mM glucose values within the same serum group; P = .05).

Fluorescence microscopy images indicating the viability of BMSCs in two-dimensional culture after the second day (increasing glucose concentrations from left to right; increasing serum concentrations from top to bottom). Calcein-AM indicates living cells by emitting green light, while ethidium bromide indicates dead cells by emitting red light. The 0.5 mM and 1.0 mM environments resulted in low viability and sparse cell distribution. The 5.0 mM and 25.0 mM environments resulted in high viability and the maintenance of high confluency. Varying the serum concentration did not produce a noticeable difference in the viability. This result was consistent throughout the course of the week (20x objective; scale bar represents 250 μm).