Biology subjects

Stout, A. J.

Publications and source records attributed to Stout, A. J..

3 recordsLinked to original sources

bioRxiv2023

Engineered autocrine signaling eliminates muscle cell FGF2 requirements for cultured meat production

Cultured meat is a promising technology that faces substantial cost barriers which are currently driven largely by the price of media components. Growth factors such as fibroblast growth factor 2 (FGF2) drive the cost of serum-free media for relevant cells including muscle satellite cells. Here, we engineered immortalized bovine satellite cells (iBSCs) for inducible expression of FGF2 and/or mutated RasG12V in order to overcome media growth factor requirements through autocrine signaling. Engineered cells were able to proliferate over multiple passages in FGF2-free medium, thereby eliminating the need for this costly component. Additionally, cells maintained their myogenicity, albeit with reduced differentiation capacity. Ultimately, this offers a proof-of-principle for lower-cost cultured meat production through cell line engineering.

bioengineering↗

bioRxiv2022

Immortalized bovine satellite cells for cultured meat applications

For cultured meat to succeed at scale, muscle cells from food-relevant species must be expanded in vitro in a rapid and reliable manner to produce millions of metric tons of biomass annually. Toward this goal, genetically immortalized cells offer substantial benefits over primary cells, including rapid growth, escape from cellular senescence, and consistent starting cell populations for production. Here, we develop genetically immortalized bovine satellite cells (iBSCs) via constitutive expression of bovine Telomerase reverse transcriptase (TERT) and Cyclin-dependent kinase 4 (CDK4). These cells achieve over 120 doublings at the time of publication and maintain their capacity for myogenic differentiation. They therefore offer a valuable tool to the field, enabling further research and development to advance cultured meat.

cell biology↗

bioRxiv2022

A Beefy-R culture medium: replacing albumin with rapeseed protein isolates

The development of cost-effective serum-free media is essential for the economic viability of cultured meat. A key challenge facing this goal is high-cost recombinant albumin that is necessary in some available serum-free media formulations. As such, there is substantial interest in finding albumin alternatives which are low-cost, effective, scalable, sustainable, and suitable for food applications. Recently, a serum-free medium termed Beefy-9 was developed for bovine satellite cells (BSCs), which relied on recombinant albumin as a key component to replace fetal bovine serum. Here we alter Beefy-9 by replacing albumin with rapeseed protein isolate, a bulk-protein solution obtained from agricultural waste-streams through simple isoelectric protein precipitation. This new medium, termed Beefy-R, improves BSC growth compared with Beefy-9 while maintaining cell phenotype and myogenicity. These results offer an effective, low-cost, and sustainable alternative to albumin for serum-free culture of muscle stem cells, thereby addressing a key hurdle facing cultured meat production.

bioengineering↗