Wharton's Jelly multipotent mesenchymal stromal cells (WJ-MSCs) hold potential for regenerative medicine, particularly in soft tissue engineering. However, their adipogenic differentiation capacity is inferior to adipose tissue-derived MSCs (AT-MSCs). This study aimed to optimize adipogenic differentiation for WJ-MSCs by leveraging insights from the comparative analysis of WJ- and AT-MSC lipidomic profiles. Lipidomic profiles of non-induced cells were compared, and adipogenic differentiation was induced with and without exogenous oleic or linoleic acid supplementation. Differentiation efficiency was determined based on lipid droplet formation, triglyceride (TG) content quantification, and the expression of adipogenic markers. Significant differences in TG composition were observed, with WJ-MSCs showing higher levels of 52-carbon TGs and AT-MSCs having more 56-carbon species. Both cell types had similar fatty acid (FA) profiles, with 18-carbon FAs making up over 50%. Adding oleic acid to the differentiation medium significantly enhanced lipid droplet formation and upregulated adipogenic markers in WJ-MSCs, aligning their adipogenic capacity more closely with AT-MSCs. In contrast, linoleic acid showed no significant benefits. The study underscores the critical role of the initial lipidomic profile in the adipogenic differentiation of MSCs. Supplementation with oleic acid represents a promising approach for improving adipogenic differentiation of WJ-MSCs and their utility in soft tissue engineering.

Charles University Prague Czech Republic

Department of Cryobiochemistry Institute of Cryobiology and Cryomedicine National Academy of Science of Ukraine Kharkiv Ukraine

Department of Neuroregeneration Institute of Experimental Medicine Czech Academy of Sciences Prague Czech Republic

Laboratory of Biomaterials and Tissue Engineering Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic

Laboratory of Mitochondrial Physiology Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic

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