A Streamlined Workflow for the Isolation of Bovine Satellite Cells From Muscle Tissue.

BACKGROUND: Satellite cells are the primary drivers of skeletal muscle growth and regeneration and are a major cell source in both muscle tissue engineering and cultivated meat. However, current bovine satellite cell isolation can be technically demanding or yield heterogeneous cell populations. METHODS: This study presents a workflow optimized for simplicity and cost-efficiency by integrating rapid muscle processing using electric mincing with magnetic-activated cell sorting targeting Integrin-α7 to reliably enrich satellite cells. The isolated cells were characterized by Pax7 immunostaining and myogenic differentiation assays. RESULTS: ITGA7-based magnetic sorting yielded an enriched satellite cell population corresponding to 49.39 ± 14.70 × 10³ cells per gram of minced tissue. The isolated cells exhibited high Pax7 expression (98 ± 1.34% Pax7-positive cells) and strong myogenic differentiation capacity (differentiation index: 94.39 ± 3.58%), confirming their identity and functional potential. CONCLUSION: By minimizing hands-on effort and avoiding specialized equipment, this workflow lowers technical and financial barriers while enabling the acquisition of highly enriched, early-passage satellite cell populations. This workflow provides an adaptable foundation for advancing cultivated meat bioprocess development and accelerating research in skeletal muscle biology.