# Optimizing chicken muscle stem cell isolation using recombinant proteolytic enzymes for cultured meat production

## TL;DR

This paper shows that using recombinant enzymes improves the isolation of chicken muscle stem cells, which is important for cultured meat production.

## Contribution

The study introduces recombinant proteolytic enzymes as a safer and more efficient method for isolating muscle stem cells compared to traditional crude enzymes.

## Key findings

- Recombinant enzymes significantly improved tissue dissociation and increased the yield of muscle stem cells.
- MSCs isolated with recombinant enzymes showed better proliferation and differentiation characteristics.
- The method offers a safer and more efficient alternative for stem cell isolation in cultured meat production.

## Abstract

Muscle satellite cells, also known as muscle stem cells (MSCs), are crucial for muscle growth and regeneration, and are among the most commonly used cell types for cultured meat production. Efficient isolation of high-quality MSCs is essential for both basic research and for the development of therapies targeting muscle-related diseases and for advancing cultured meat production. Traditional isolation methods often use crude proteolytic enzymes, which can lead to contamination and cytotoxicity. This study aimed to improve the isolation of muscle stem cells by using recombinant proteolytic enzymes—collagenase and thermolysin—while comparing their performance to crude enzyme treatments. The isolation of MSCs from chicken muscle tissue was conducted using both crude pronase and recombinant enzymes. The effectiveness of each method was evaluated based on tissue dissociation efficiency and the quantity of MSCs isolated. The results showed that the combination of recombinant proteolytic enzymes significantly improved muscle tissue dissociation, resulting in a higher yield of MSCs compared to crude enzymes. Furthermore, the MSCs isolated using recombinant enzymes exhibited comparable or superior proliferation and differentiation characteristics, including myogenic gene and protein expression and creatine kinase activity. These findings suggest that recombinant proteolytic enzymes offer a safer and more efficient alternative to crude enzymes for isolating MSCs. This method has potential applications in muscle research and the production of cultured meat, improving both safety and efficiency in stem cell-based technologies and fostering advancements in these fields.

The online version contains supplementary material available at 10.1007/s44463-025-00051-8.