Mitochondrial function determines the viability and osteogenic potency of human mesenchymal stem cells

Area of Science:

• Cell Biology
• Biotechnology
• Regenerative Medicine

Background:

• Advanced therapies medicinal products (ATMPs) require robust characterization for safety and efficacy.
• Mitochondrial function is increasingly recognized as critical for human mesenchymal stem cell (hMSC) differentiation.
• Current regulatory methods may not fully capture the nuances of cell quality for ATMPs.

Purpose of the Study:

• To investigate the viability and osteogenic differentiation potential of hMSCs for ATMP applications.
• To analyze the mitochondrial inner membrane potential (DeltaPsi(m)) as a key indicator of hMSC quality.
• To establish a correlation between DeltaPsi(m) and osteogenic differentiation capacity.

Main Methods:

• Flow cytometry was used to measure DeltaPsi(m) with JC-1 dye.
• Propidium iodide and AnnexinV staining assessed plasma membrane integrity and phosphatidylserine organization.
• Menadione induced apoptosis; MTT assays, alkaline phosphatase activity, and calcium deposition measured osteogenic differentiation.

Main Results:

• Cells were categorized into four quality groups based on JC-1 fluorescence, morphology, and membrane integrity.
• A significant correlation was demonstrated between JC-1 fluorescence (DeltaPsi(m)) and osteogenic differentiation potential.
• JC-1 analysis proved effective in assessing both hMSC viability and predicting differentiation capacity.

Conclusions:

• Mitochondrial inner membrane potential (DeltaPsi(m)) is a reliable metric for evaluating hMSC quality for ATMPs.
• JC-1 staining offers a dual assessment of cell viability and osteogenic differentiation potential.
• This analytical approach enhances the characterization of hMSCs, improving the safety and predictability of ATMPs.