stemprotm adipogenesis differentiation kit  (Thermo Fisher)

Journal: International Journal of Molecular Medicine

Article Title: Nervonic acid improves fat transplantation by promoting adipogenesis and angiogenesis

doi: 10.3892/ijmm.2024.5432

Figure Lengend Snippet: Effect of NA on adipogenesis. (A) Cell viability of AD-MSCs as measured by O.D. values at day 3 and 7 after treatment with DMSO or serial doses of NA. (B) Representative images of AD-MSCs after 72 h of NA treatment. Yellow arrows indicate the oil droplets formed after NA treatment in the normal proliferation condition. Scale bars: 100 μ m. (C) Representative images of Oil Red O staining of AD-MSCs on day 7 after adipogenic differentiation and treatment with DMSO or NA. Scale bars: 100 μ m. (D) Relative amounts of lipid accumulation after 7 days of adipogenic differentiation in cells treated with DMSO or NA. Statistical significance was assessed by one-way ANOVA: *** P<0.001. (E) Representative images of Oil Red O staining of AD-MSCs after 7, 14 and 21 days of adipogenic differentiation and treatment with DMSO or 160 μ M NA. Scale bars: 100 μ m. (F and G) Relative amounts of lipid accumulation after 7, 14 and 21 days of adipogenic differentiation and treatment with DMSO or 160 μ M NA. (F) Increase in lipid accumulation by differentiation period and NA treatment. Relative lipid accumulation was normalized to the DMSO group at day 7. Statistical significance was assessed by one-way ANOVA: *** P<0.001. (G) Change in lipid accumulation at each differentiation timepoint (7, 14 and 21 days) in the NA group versus the DMSO group. Relative lipid accumulation was normalized to the DMSO group at each time point. Statistical significance was assessed by unpaired Student's t-test: *** P<0.001. (H) Reverse transcription-quantitative PCR analysis of adipogenic markers after 7, 14 and 21 days of adipogenic differentiation and treatment with DMSO or 160 μ M NA. Statistical significance was assessed by unpaired Student's t-test: * P<0.05, ** P<0.01 and *** P<0.001. Data are presented as the mean ± SD. AD-MSCs, adipose-derived mesenchymal stem cells; DMSO, dimethyl sulfoxide; LPL , lipoprotein lipase; NA, nervonic acid; O.D., optical density.

Article Snippet: For adipogenic differentiation, MSCs were cultured in an adipogenesis differentiation medium (StemProTM Adipogenesis Differentiation kit; cat. no. A1007001; Gibco; Thermo Fisher Scientific, Inc.) containing different concentrations of NA (0, 40, 80, 120, 160, 200, 240 and 280 μ M), according to the manufacturer's instructions.

Techniques: Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Derivative Assay

Journal: International Journal of Molecular Medicine

Article Title: Nervonic acid improves fat transplantation by promoting adipogenesis and angiogenesis

doi: 10.3892/ijmm.2024.5432

Figure Lengend Snippet: Signaling pathways regulating adipogenesis are affected by NA. (A) Western blot analysis of adipogenic markers after 14 days of adipogenic differentiation and treatment with DMSO or 160 μ M NA. (B) Western blot analysis of Akt and mTOR phosphorylation. (C) Western blot analysis of GSK3β expression and β-catenin phosphorylation. (D) Western blot analysis of Smad1/5 and ERK1/2 phosphorylation. (E) Graphical representation of signaling pathways affected by NA during adipogenesis. Created with BioRender.com . Statistical significance was assessed using unpaired Student's t-test: * P<0.05, ** P<0.01 and *** P<0.001. Data are presented as the mean ± SD. DMSO, dimethyl sulfoxide; LPL, lipoprotein lipase; NA, nervonic acid; p-, phosphorylated.

Article Snippet: For adipogenic differentiation, MSCs were cultured in an adipogenesis differentiation medium (StemProTM Adipogenesis Differentiation kit; cat. no. A1007001; Gibco; Thermo Fisher Scientific, Inc.) containing different concentrations of NA (0, 40, 80, 120, 160, 200, 240 and 280 μ M), according to the manufacturer's instructions.

Techniques: Protein-Protein interactions, Western Blot, Phospho-proteomics, Expressing

Journal: International Journal of Molecular Medicine

Article Title: Nervonic acid improves fat transplantation by promoting adipogenesis and angiogenesis

doi: 10.3892/ijmm.2024.5432

Figure Lengend Snippet: Transcriptomic changes in differentiating AD-MSCs after NA treatment. (A) Principal component analysis of AD-MSCs treated with DMSO or NA during adipogenesis. (B) Gene Ontology analysis of the biological processes associated with differentially expressed genes in the NA-treated group against the DMSO-treated group at day 7 of adipogenesis. (C) Heatmap of the expression of genes involved in 'mTOR signaling, 'positive regulation of mTOR signaling' (left) and 'adipose tissue development' (right) in the NA-treated group versus the DMSO-treated group. (D) Heatmap of the expression of genes involved in 'angiogenesis' (left) and 'positive regulation of vascular endothelial growth factor production' (right) in the NA-treated group versus the DMSO-treated group. AD-MSCs, adipose-derived mesenchymal stem cells; DMSO, dimethyl sulfoxide; NA, nervonic acid.

Article Snippet: For adipogenic differentiation, MSCs were cultured in an adipogenesis differentiation medium (StemProTM Adipogenesis Differentiation kit; cat. no. A1007001; Gibco; Thermo Fisher Scientific, Inc.) containing different concentrations of NA (0, 40, 80, 120, 160, 200, 240 and 280 μ M), according to the manufacturer's instructions.

Techniques: Expressing, Derivative Assay