A Multi-Institution Biobanking Pipeline for Primary Human Satellite Cells and Fibro-Adipogenic Progenitors

Area of Science:

• Muscle biology
• Regenerative medicine
• Cell biobanking

Background:

• Satellite Cells (SCs) and Fibro-Adipogenic Progenitors (FAPs) are vital for skeletal muscle homeostasis and repair.
• Challenges in co-isolation and lack of standardized biobanking limit translational research using primary human SCs and FAPs.

Purpose of the Study:

• To establish a comprehensive pipeline for cryopreservation, transport, and utilization of human SCs and FAPs.
• To ensure the viability, phenotype, and differentiation capacity of these cells post-biobanking.
• To facilitate multi-site collaboration and broaden access to human primary muscle cells for research.

Main Methods:

• Development of a pipeline for cryopreservation and cold-chain transport of co-isolated human SCs and FAPs.
• Assessment of lineage-specific marker retention (Pax7, MyoD, CD56 for SCs; PDGFRα, TE7 for FAPs).
• Evaluation of myogenic differentiation capacity for SCs and fibrogenic/adipogenic capacity for FAPs post-transport.
• Incorporation of biobanked SCs into 3D in vitro muscle constructs.

Main Results:

• Cells maintained lineage-specific markers and pre-biobanking phenotype after the pipeline process.
• SCs retained robust myogenic differentiation capacity.
• FAPs retained fibrogenic and adipogenic differentiation capacity.
• Biobanked SCs were successfully used in 3D muscle constructs for in vitro studies.

Conclusions:

• The established pipeline enables reliable biobanking and transport of human SCs and FAPs, preserving their key characteristics.
• This framework supports multi-site collaborations, enhancing the scalability and translatability of human-based New Approach Methodologies (NAMs) in skeletal muscle research.