Comparison of proliferation, differentiation and migration ability of adipose-derived stem cells in three different granular fats

Objective To study the effects of three different granular fats on proliferation, differentiation and migration of adipose-derived stem cells. Methods Ten patients were selected for lumbar liposuction. The adipose tissue was obtained with different sized side-hole fat aspiration devices: 10 ml of Macrofat (n=10), 10 ml of Microfat (n=10) and 10 ml of Nanofat (n=10). Enzyme digestion method was used to separate and extract adipose-derived stem cells(ADSCs). The differences of vascular matrix components in these 3 kinds of fat sources were analyzed. XTT assay was used to detect proliferation and growth ability. The migration ability of the cell injury model was observed in vitro, and the multi-directional differentiation ability was compared by the analysis of adipogenic and osteogenic induction. The experimental data were analyzed using SPSS 13.0 software. One-Way ANOVA was used to compare the difference of multiple groups. P<0.05 was considered as statistically significant. Results The detection of adipose-derived stem cells was by flow cytometry and it showed Macrofat, Microfat and Nanofat was(4.23±0.37)×105, (2.29±0.33)×105 and(1.56±0.16)×105. The content of fat-derived stem cells in Macrofat was the highest, followed by Microfat, and the content of Nanofat was the least (F=209.533, P<0.001). XTT assay showed that the proliferative ability of adipose-derived stem cells in the first two generations of Macrofat was the highest, followed by Microfat, while the proliferation of Nanofat was the lowest (the absorbance in 3 groups increased in a time-dependent manner). There was no significant difference in the proliferative capacity of the third generation of 3 groups (the absorbance of these 3 groups were compared at each time point). The cellular trauma model showed that the first generation of Macrofat-derived stem cells had the best migration ability, followed by Microfat, while the Nanofat had the weakest migration ability(Compared with the remaining area of wounds at 0 h, 12 h, 24 h point between the three groups, F=306.370, 1409.907, P<0.001). From the second generation, the migration ability of each group of ADSCs gradually self-repaired, and the migration ability at 12 h was similar to that of the first generation (F=11.410, P<0.001), but there was no significant difference in 24 h (F=0.070, P=0.933). Oil red O and alizarin red staining showed that the first generation of Macrofat and Microfat had better fat-forming and osteogenic differentiation ability than Nanofat(F=523.532, 620.022, P<0.001). However, there was no significant change after the second generation (F=2.144, 0.866, P=0.137, 0.432). Conclusions In the process of adipose tissue extraction of Nanofat, the production and activity of adipose-derived stem cells was impacted. However, in the process of culture and passage, the cell activity, proliferation ability, migration ability and differentiation ability can be achieved through self-repair, evenclose to the level of Macrofat or Microfat. Key words: Granular fat; Nanofat; Adipose-derived stem cells; Proliferation; Differentiation; Migration