Simple and effective serum-free medium for sustained expansion of bovine satellite cells for cell cultured meat

Abstract

Cell-cultured meat offers the potential for a more sustainable, ethical, resilient, and healthy food system. However, research and development has been hindered by the lack of suitable serum-free media that enable the robust expansion of relevant cells (e.g., muscle satellite cells) over multiple passages. Recently, a low-cost serum-free media (B8) was described for induced pluripotent stem cells. Here, we adapt this media for bovine satellite cells and show that the addition of a single component, recombinant albumin, renders B8 suitable for the long-term expansion of cells without sacrificing myogenicity. We show that this new media (Beefy-9) maintains robust cell growth over the entire culture period tested (seven passages) with an average growth rate of 39 hours per population doubling). Along with demonstrated efficacy for bovine cells, this work provides a promising starting-point for developing serum-free media for cultures from other meat-relevant species. Ultimately, this work offers a promising foundation for escaping the reliance on serum in cultured meat research, thereby accelerating the field.

Review of the Current Research on Fetal Bovine Serum and the Development of Cultured Meat

“…Although the initial cell adhesion efficiency and cell viability were low during cell culture using a serum-free medium, the cell growth efficiency was improved to a value close to that of the serum-containing medium through partial medium replacement and removal of antibiotics. In another study, Stout et al (2021) found that cell activity was maintained by subculturing bovine satellite cells in a serum-free medium and evaluated the possibility of reducing the medium cost by predicting the optimal amount of medium and additives. In particular, long-term cell expansion was possible when recombinant albumin was added to the developed Beefy-9 medium.…”

Scaffolding Biomaterials for 3D Cultivated Meat: Prospects and Challenges

“…Although substantial progress has been achieved in recent years by both for‐profit companies and academic groups, CM remains a relatively new area of investigation. As might be expected for a nascent field that draws heavily on knowledge gained from previous work aimed at a distinct set of problems, review, and perspective papers about CM (at least 50 by our estimate) [ 10–14,16,18,23,28,37,44,64–109 ] currently outnumber original research papers where CM production is the primary intended application (at least 24) [ 15,59,110–131 ] or CM‐related patents (at least 24). [ 132–155 ] These estimates do not include LCA or TEA studies, consumer acceptance studies, or purely computational analyses.…”

Cell Sources for Cultivated Meat: Applications and Considerations throughout the Production Workflow

“…As a result, replacing FBS with serum-free supplements or chemically defined serum alternatives is desirable to eliminate animal-derived components from culture and to create media that have consistent performance. In recent years, the development of robust xeno-free medium formulations such as Essential 8™, TeSR™, and FBM™ has enabled the removal of serum from cell culture [85][86][87]. However, further advancements are still needed to create chemically defined media formulations that are consistently as effective as serum-based media in promoting cell growth, particularly for livestock animal cells [86,88,89].…”