Effects of Macromolecular Crowding on Human Adipose Stem Cell Culture in Fetal Bovine Serum, Human Serum, and Defined Xeno-Free/Serum-Free Conditions

Figure 3

Morphology, cell number, and metabolic activity of ASCs under different culture conditions. (a) ASCs cultured under MMC adopted rounded morphology and became larger in size. The cell number under MMC in XF/SF cultures significantly decreased compared with that in standard cultures. Passage = 3. Scale bar 500 μm (100 μm in XF/SF inset). (b) The DNA amount indicating cell number in different culture conditions was analyzed with CyQUANT cell proliferation assay. Fluorescence signals were measured at 480/520 nm. A significantly higher cell number was observed in standard HS media at day 1 and in standard FBS media at days 4 and 7 and compared with that in MMC cultures. Under MMC, at time points—day 7 and 11—the cell number was significantly higher in HS media compared with that in FBS media. A significantly higher cell number was observed under standard cultures in XF/SF conditions compared with that in standard FBS and HS cultures at each time point—days 1, 4, 7, and 11. (c) The metabolic activity of ASCs was studied with CCK-8 assay and normalized to total DNA quantified by CyQUANT cell proliferation assay. Fluorescence signals were measured at 450 nm. A statistically significant increase in metabolic activity in FBS media under MMC was observed at day 1 compared with that in standard FBS media. ASCs in FBS media under MMC had a significantly higher metabolic activity compared with cells in HS media under MMC at days 4, 7, and 11 time points. Although a decreased cell number and metabolic activity was observed in XF/SF medium under MMC, no statistical differences could be established due to the low number donors that survived under MMC. indicates ; indicates . Data are presented as mean ± SD. FBS, fetal bovine serum; HS, human serum; XF/SF, xeno-free/serum-free; MMC, macromolecular crowding.

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