Data supporting the article "ENHANCING ADIPOGENESIS IN WHARTON'S JELLY MULTIPOTENT MESENCHYMAL STROMAL CELLS: THE ROLE OF OLEIC AND LINOLEIC ACID SUPPLEMENTATION"

Data supporting the article "ENHANCING ADIPOGENESIS IN WHARTON'S JELLY MULTIPOTENT MESENCHYMAL STROMAL CELLS: THE ROLE OF OLEIC AND LINOLEIC ACID SUPPLEMENTATION"

Figure 1. Adipogenic differentiation of WJ-MSCs and AT-MSCs (N=4 for each) following the standard induction protocol:

A – Lipid droplets in control and induced cultures stained by Nile Red (21 days of culture);

B – TG accumulation in control groups and after directed adipogenic differentiation (21 days of culture). Data is presented as Mean ± SD and shown in duplicates. Note: * - p<0.05; ** - p<0.01; *** - p<0.001; **** - p<0.0001.

 Figure 2. Lipidomic profiles of WJ-MSCs and AT-MSCs (N=4) based on liquid chromatography-mass spectrometry (LC-MS). Abbreviations: phosphatidylethanolamines (PE), phosphatidylinositols (PI), phosphatidylserines (PS), cholesterol esters (CE), lysophosphatidylcholines (LPC), lysophosphatidylethanolamines (LPE), free fatty acids (FAs), and triglycerides (TG). Data is presented as Mean ± SD. Signal intensities were normalized to the respective total ion count (TIC) before subsequent statistical analysis. Note: * - p<0.05.

Figure 3. Comparison of TG proportion in WJ-MSCs and AT-MSCs (N=4): A – Distribution of different TG species in WJ-MSCs and AT-MSCs; B, C - Distribution of TGs by carbon number and number of double bonds in WJ-MSCs (B) and AT-MSCs (C) (N=4). The heatmap plot represents the proportion of different TG species among all TGs. The black frame highlights differences between cell types, particularly in long-chain TGs (54–58 carbons), where AT-MSCs shift towards a higher number of double bonds relative to WJ-MSCs.

Figure 4. Comparison of FA proportion in WJ-MSCs and AT-MSCs (N=4): A – Differences in FA profile in WJ-MSCs and AT-MSCs; B – Share of 18-carbon fatty acids, namely oleic acid, linoleic and linolenic acids, among all FA; Data is presented as Mean ± SD. Note: * - p<0.05.

 Figure 5. Adipogenic differentiation of WJ-MSCs and AT-MSCs (N=4) following OA (100 µM), LA (100 µM), or their mixture (50 µM each) supplementation. A - Lipid droplets after 21 days of induction by standard differentiation media and with FA addition, Nile red-stained cultures. B - TG accumulation in WJ-MSCs following FA supplementation; C - TG accumulation in AT-MSCs following FA supplementation. Note: * - p<0.05; ** - p<0.01

 Figure 6. Expression of key adipogenic markers in WJ-MSCs after differentiation with and without exogenous FA supplementation (N=4). Data is presented as Mean ± SD of Log2-fold change values vs non-differentiated control cultures assessed by RT-PCR. Note: * - p<0.05.

 Figure 7. Expression of key adipogenic markers in AT-MSCs after differentiation with and without exogenous FA supplementation (N=4). Data is presented as Mean ± SD of Log2-fold change values vs non-differentiated control cultures assessed by RT-PCR. Note: * - p<0.05.

Table 1. Lipidomic data that shows the differences between WJ-MSCs and AT-MSCs.Signal intensities were normalized to the respective total ion count (TIC) before subsequent statistical analysis.

Photo 1. Lipid droplets in control WJ-MSCs cultures stained by Nile Red (21 days of culture);

Photo 2. Lipid droplets in WJ-MSCs cultures after standard adipogenic induction stained by Nile Red (21 days of culture);

Photo 3. Lipid droplets in WJ-MSCs cultures after adipogenic induction and supplementation with OA stained by Nile Red (21 days of culture);

Photo 4. Lipid droplets in WJ-MSCs cultures after adipogenic induction and supplementation with LA stained by Nile Red (21 days of culture);

Photo 5. Lipid droplets in WJ-MSCs cultures after adipogenic induction and supplementation with LA and OA stained by Nile Red (21 days of culture);

Photo 6. Lipid droplets in control AT-MSCs cultures stained by Nile Red (21 days of culture);

Photo 7. Lipid droplets in AT-MSCs cultures after standard adipogenic induction stained by Nile Red (21 days of culture);

Photo 8. Lipid droplets in AT-MSCs cultures after adipogenic induction and supplementation with OA stained by Nile Red (21 days of culture);

Photo 9. Lipid droplets in AT-MSCs cultures after adipogenic induction and supplementation with LA stained by Nile Red (21 days of culture);

Photo 10. Lipid droplets in AT-MSCs cultures after adipogenic induction and supplementation with LA and OA stained by Nile Red (21 days of culture);