Automated analysis of mitochondrial dimensions in mesenchymal stem cells: Current methods and future perspectives

Abstract

As centre of energy production and key regulators of metabolic and cellular signaling pathways, the integrity of mitochondria is essential for mesenchymal stem cell function in tissue regeneration. Alterations in the size, shape and structural organization of mitochondria are correlated with the physiological state of the cell and its environment and could be used as diagnostic biomarkers. Therefore, high-throughput experimental and computational techniques are crucial to ensure adequate correlations between mitochondrial function and disease phenotypes. The emerge of microfluidic technologies can address the shortcomings of traditional methods to determine mitochondrial dimensions for diagnostic and therapeutic use. This review discusses optical detection methods compatible with microfluidics to measure mitochondrial dynamics and their potential for clinical stem cell research targeting mitochondrial dysfunction.

Keywords: Automated mitochondrial analysis; MSC-Based therapies; Mesenchymal stem cells; Microfluidic systems; Mitochondrial dynamics.

Fig. 1

Processes that influence the homeostasis of mitochondrial dimensions.

Fig. 2

Effect of actin-based cytoskeleton disruption on mitochondrial distribution. Control MSCs show a normal mitochondrial distribution (red) with minimal perinuclear accumulation (nucleus in blue). Treatment of MSCs for 24 h with 16 μM cytochalasin B (24 h CB) triggers actin filament depolymerization, which results in increased perinuclear accumulation of mitochondria.

Fig. 3

Mitochondrial transport through nanotunnels by MSCs. Mitochondrial nanotunnels (MNTs, green) are tubular structures containing mostly of F-actin that can be formed by MSCs to transport mitochondrial particles or other cellular components (e.g., extracellular vesicles) to other MSCs or other cell types (e.g., endothelial cells, platelets) to restore their vitality and support their function, preventing apoptosis.

Fig. 4

Overview of microfluidic technologies. Lab-on-a-chip, organ-on-a-chip and microphysiological systems A) with multi-cellular 2D and 3D environment to investigate cellular function, interaction, and tissue context. B) Oxygen and glucose consumption as well as ATP and pH-value measures can be determined using microfluidic devices.