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

Figure 5

Quantitative Nile red staining indicating the adipogenic differentiation and collagen IV staining indicating adipogenic matrix deposition under different conditions. (a) The adipogenic differentiation was analyzed using Nile red staining that was normalized to cell number (b). In FBS and HS conditions, ASCs accumulated significantly more lipid content under induction with MMC compared with those under standard induction. A stronger differentiation was observed in HS media compared with that in FBS media in each induction group and significantly more lipid droplets were observed in standard HS induction media compared with those in standard FBS induction. Scale bar 500 μm; indicates ; indicates . Data are presented as mean ± SD. (c) Matrix deposition was evaluated using Col IV staining after adipogenic induction. Col IV deposition was enhanced under MMC induction compared with induction under MMC-free media in both FBS and HS conditions. Additionally, Col IV was more enhanced in HS media compared with that in FBS media based on qualitative analysis. Col IV deposition was in line with the results of lipid droplet accumulation that was confirmed using Nile red staining. Scale bar 500 μm. FBS, fetal bovine serum; HS, human serum; E ± MMC, expansion under macromolecular crowding/in standard medium; D ± MMC, differentiation under macromolecular crowding/in standard medium.

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