Lipogems Product Treatment Increases the Proliferation Rate of Human Tendon Stem Cells without Affecting Their Stemness and Differentiation Capability

Figure 4

Effects of the Lipogems product on tendon marker expression. (a) Gene expression of TNMD, Tenascin C, and COL1A1 by Real-Time PCR at 96 h of treatment with the Lipogems product. Values are expressed as fold-changes relative to control cells. Effects of the Lipogems product on the in vitro differentiation of hTSCs toward the adipogenic and the osteogenic phenotypes. Adipogenic and osteogenic differentiation ability of hTSCs cultured with the Lipogems product in the appropriate differentiation medium was evaluated by Oil Red O (b) and Alizarin Red-S (c) staining, respectively. (b) Lipid intracellular droplets (red) in the adipocytes were stained with Oil Red O solution. (c) Alizarin Red-S staining revealed the presence of calcium deposits (yellowish-brown). Typical results are shown. Original magnification ×10. Effect of Lipogems treatment on stem cells marker (Nanog, Oct4, and KLF4) (d) and VEGF (e) expression by Real-Time PCR at 96 h of treatment with the Lipogems product. Values are expressed as fold-changes relative to control cells. Effect of the Lipogems product on hTSCs migration. (f) Representative time-lapse migration images of control and Lipogems-treated cells. Images were acquired at 0 and 25 h in in vitro wound-healing assay. Original magnification ×5. The migration rate was measured by quantifying the total area of the wounded region lacking cells. The average percentages of recovered area obtained from three different experiments at 17, 20, 25, 30, and 42 h of treatment with the Lipogems product, as compared to control cells. All quantitative data are expressed as the means ± SD of three different experiments. values were calculated using Student’s -test. Only values < 0.05 are indicated: , compared to control cells.

(a) Tendon marker expression

(b) Adipogenic differentiation

(c) Osteogenic differentiation

(d) Stem cell marker expression

(e) VEGF expression

(f) Wound-healing assay