Primary culture of myogenic progenitor cells derived from embryologically diverse post-mortem human skeletal muscles

Primary human satellite cells (SCs) are essential for skeletal muscle regeneration research, but access to tissue is constrained by the need for invasive biopsies. Post-mortem muscle represents a potential alternative, yet data on establishing expandable myogenic cultures from multiple human muscle groups are sparse.

To develop a workflow for isolating, expanding, and morphologically assessing myogenic progenitor cells from embryologically distinct muscles of human body donors.

Eighteen biopsies from six muscles (erector spinae, vastus lateralis, brachioradialis, masseter, extraocular muscles, diaphragm) were collected from three body donors (age 79-85 years; post-mortem intervals 13-30 h). Cells were isolated by enzymatic dissociation, enriched by fluorescence-activated cell sorting (CD31⁻/CD45⁻/CD56⁺), and cultured for six weeks. Differentiation was assessed by phase-contrast microscopy.

Viable cells were isolated from all donors (median viability 10.3%). Cultures were established from one of three donors; the remaining two failed due to insufficient biopsy size (procedural) and low viability despite the shortest post-mortem interval (biological), respectively. In the successful donor, five of six muscles including somitic and non-somitic origins gave rise to adherent cultures forming structures morphologically consistent with myotubes. Culture viability increased from 11.6% to 36.7% over six weeks.

Post-mortem skeletal muscle from anatomical body donors can yield cells capable of in vitro expansion and morphological myogenic differentiation across embryologically diverse muscle groups. Inter-donor variability limits consistent success. This pilot study provides a practical workflow and documents both successful and unsuccessful outcomes to inform future optimization.