Biology subjects

Hueber, R.

Publications and source records attributed to Hueber, R..

3 recordsLinked to original sources

bioRxiv2025

Optimisation of skeletal muscle sampling for cultured fat production

To produce cultured meat at a commercially-relevant scale, bioprocesses must be robust, standardised and cost-efficient. In this respect, the limited in vitro lifespan of primary cells poses a major challenge, which requires regular cell sourcing by means of biopsy. We have previously shown that muscle-derived fibro-adipogenic progenitor cells (FAPs) represent a promising starting cell type for cultured fat production. These cells proliferate for a high number of population doublings (PDs) in serum-free growth medium. However, with accumulating PDs, FAPs lose their adipogenic differentiation capacity, thereby limiting the amount of cultured fat that can be produced from a given starting sample. Donor animal characteristics, including age, may affect this loss of differentiation. Here, we performed a longitudinal biopsy study to ask whether physiological differences between donor animals are reflected in FAP cell biology, and if this has an effect on loss of FAP differentiation capacity. We sampled twelve Limousin cattle over the first two years of their lives, successfully performing 144 muscle biopsies. Animals younger than six months demonstrated higher FAP yields per gram of muscle tissue, making them ideal donors for cultured fat production, but we observed little difference in the proliferation or differentiation of derived cell cultures. This work also provides valuable insights into the tissue harvesting process, highlighting the need for new, robust and standardised biopsy systems to mitigate contamination risk.

cell biology↗

bioRxiv2022

Single-cell analysis of bovine muscle-derived cell types for cultured meat production

Cultured meat technologies leverage the proliferation and differentiation of animal-derived stem cells ex vivo to produce edible tissues for human consumption in a sustainable fashion. However, skeletal muscle is a dynamic and highly complex tissue, involving the interplay of numerous mono- and multinucleated cells, including muscle fibres, satellite cells (SCs) and fibro-adipogenic progenitors (FAPs), and recreation of the tissue in vitro thus requires the characterisation and manipulation of a broad range of cell types. Here, we use a single-cell RNA sequencing approach to characterise cellular heterogeneity within bovine muscle and muscle-derived cell cultures over time. Using this data, we identify numerous distinct cell type, and develop robust protocols for the easy purification and proliferation of several of these populations. We note overgrowth of undesirable cell types within heterogeneous proliferative cultures as a barrier to efficient cultured meat production, and use transcriptomics to identify conditions that favour the growth of SCs in the context of serum-free medium. Combining RNA velocities computed in silico with time-resolved flow cytometric analysis, we characterise dynamic subpopulations and transitions between active, quiescent, and committed states of SCs, and demonstrate methods for modulation of these states during long-term proliferative cultures. This work provides an important reference for advancing our knowledge of bovine skeletal muscle biology, and its application in the development of cultured meat technologies.

cell biology↗

bioRxiv2022

Comparative analysis of cattle breeds as satellite cells donors for cultured beef

BackgroundCultured meat is a promising new field with the potential for considerable environmental and animal welfare benefits. One technological approach to cultured meat production utilises the proliferative and differentiative capacity of muscle-derived satellite cells (SCs) to produce large volumes of cultured muscle tissue from small biopsies of donor animals. Differing genotypes between cattle breeds lead to predictable phenotypic traits, resulting in breeds being favoured for their respective meat or milk production characteristics in the livestock industry. However, whilst these breeds show significant differences in muscle growth, it is unclear whether the physiological differences observed between them in vivo are reflected in differences in SC behaviour in vitro, particularly with respect to proliferation, differentiation and cellular longevity, and hence whether particular breeds might represent preferred SC donors for a cultured beef bioprocess. ResultsComparing SCs isolated from five breeds (Belgian Blue, Holstein Friesian, Galloway, Limousin and Simmental), we found that the proliferation rates were largely unaffected by the donor breed. In contrast, potentially meaningful differences were observed in the kinetics and extent of myogenic differentiation. Furthermore, whilst differentiation dropped for all breeds with increasing population doublings (PDs), SCs from Belgian Blue and Limousin cattle showed significantly longer retention of differentiation capacity over long-term passaging. ConclusionSCs from all breeds were able to proliferate and differentiate, although Limousin and (particularly) Belgian Blue cattle, both breeds commonly used for traditional meat production, may represent preferred donors for cultured beef production.

cell biology↗