Effect of fatty acids on mitochondrial biogenesis during mouse embryonic stem cell differentiation

Fatty acids (FAs) and their metabolites may affect and improve mitochondrial biogenesis in mouse embryonic stem cells (mESCs). Mitochondria are dynamic organelles that are responsible for adenosine triphosphate (ATP) production and are implicated in cellular functions such as proliferation, differentiation, cell cycle, reprogramming, aging, and apoptosis, with normal morphology required for cellular functions. Thus, mitochondrial morphology may change to meet the energy requirement. This study examines the impact of FAs, such as eicosapentaenoic acid (EPA) and linoleic acid (LA), on mitochondrial biogenesis through a quantitative analysis of multiple parameters. Three-day-old mESCs under differentiation conditions were treated with 50 µM FAs for 48 hours, and the results were compared with untreated and vehicle (dimethyl sulfoxide, DMSO). Mitochondrial configurations in the two-dimensional (2D) projection were determined and quantified using ImageJ. Treatment with 50 µM of FAs significantly increased the quantity of mitochondria, including mitochondrial count, area, perimeter, form factor, the number of branches, the number of branch junctions, and the length of branches, when compared to the control and vehicle groups, hence signifying improved mitochondrial interconnectivity, suggesting the enhancement of mitochondrial biogenesis. These findings explain the role of FAs in promoting mitochondrial biogenesis and suggest potential therapeutic applications for controlling metabolic diseases associated with impaired mitochondrial function.