An Optimized Pre-Plating Method for the Isolation and Culture of Human Primary Satellite Cells

Muscle satellite cells play an essential role in postnatal skeletal muscle growth, homeostasis, hypertrophy, and regeneration following injury. Efficient primary culture methods are crucial for the in vitro analysis of satellite cells to investigate the molecular mechanisms underlying the adaptation and regeneration of skeletal muscles. In this protocol, we describe a refined and accessible pre-plating method for the efficient isolation of human satellite cells that does not require specialized equipment or advanced instrumentation. Furthermore, we provide comparative data on various growth medium formulations to identify the optimal and cost-effective conditions for human satellite cell culture. Through systematic optimization of both the isolation technique and culture conditions, we established a practical and reproducible protocol specifically tailored to human skeletal muscles. This methodology enables more accurate and clinically relevant investigations into human muscle biology.