Advances in primary cell isolation, sorting, and preservation from livestock for cultured meat applications

Cultured meat production relies on the efficient: isolation, purification, expansion, and preservation of livestock-derived primary cells with robust proliferation and differentiation potential. Satellite cells and fibro-adipogenic progenitors from muscle and adipose tissues are key cellular sources for the in vitro reconstruction of meat-like structures. This review critically examines current methodologies used for primary cell processing in cultured meat scalable bioprocessing research. Tissue dissociation strategies are classified as enzymatic, non-enzymatic, and hybrid protocols and compared in terms of yield, scalability, and preservation of cell functionality. Cell purification techniques include: density gradient centrifugation, pre-plating, fluorescence-activated cell sorting (FACS), and magnetic-activated cell sorting (MACS), each offering different levels of specificity, throughput, and compatibility with food-grade applications. Long-term storage via cryopreservation is essential for establishing reliable cell banks. However, challenges remain in minimizing cryoinjury, selecting optimal cryoprotective agents, and maintaining post-thaw viability. Emerging alternatives, such as temperature-responsive cell sheet technology and biochemical protection strategies, are highlighted for their potential to improve cell recovery and functionality. Unlike existing reviews that often focus on isolated aspects, this article integrates dissociation, purification, and preservation into one framework. It highlights how optimized protocols can overcome current bottlenecks in cell processing. This review emphasizes the importance of standardized and food-safe platforms. Such advances will support commercialization and sustainable delivery of cultured meat products.