Cultivation of Bovine Mesenchymal Stem Cells on Plant-Based Scaffolds in a Macrofluidic Single-Use Bioreactor for Cultured Meat

Abstract

Reducing production costs, known as scaling, is a significant obstacle in the advancement of cultivated meat. The cultivation process hinges on several key components, e.g., cells, media, scaffolds, and bioreactors. This study demonstrates an innovative approach, departing from traditional stainless steel or glass bioreactors, by integrating food-grade plant-based scaffolds and thermoplastic film bioreactors. While thermoplastic films are commonly used for constructing fluidic systems, conventional welding methods are cost-prohibitive and lack rapid prototyping capabilities, thus inflating research and development expenses. The developed laser welding technique facilitates contamination-free and leakproof sealing of polyethylene films, enabling the efficient fabrication of macrofluidic systems with various designs and dimensions. By incorporating food-grade plant-based scaffolds, such as rice seeded with bovine mesenchymal stem cells, into these bioreactors, this study demonstrates sterile cell proliferation on scaffolds within macrofluidic systems. This approach not only reduces bioreactor prototyping and construction costs but also addresses the need for scalable solutions in both research and industrial settings. Integrating single-use bioreactors with minimal shear forces and incorporating macro carriers such as puffed rice may further enhance biomass production in a scaled-out model. The use of food-grade plant-based scaffolds aligns with sustainable practices in tissue engineering and cultured-meat production, emphasizing its suitability for diverse applications.

Cultivation of Bovine Lipid Chunks on Aloe vera Scaffolds

“…The design of either overhead aeration (as demonstrated in [42]) or with an airlift, by simply using tubes of varying lengths. Given that different scaffolds demand different agitation mechanisms and aeration techniques, had we applied the current design to puffed rice, the outcomes would have been detrimental, leading to a lack of proliferation.…”

“…This bioprocess holds promise for creating bovine fatlike chunks to serve as additives in plant-based meat alternatives. By capitalizing on existing byproducts and harnessing bioprocessing technologies, our study contributes to expediting the transition toward sustainable and efficient cultured meat production, addressing the escalating demand for protein sources while mitigating environmental impacts [8,42].…”

“…Our experimentation yielded promising findings regarding the structural integrity of the Aloe vera scaffolds during the cultivation process. While in parallel to cultivation in multiwell plates in static conditions, we have evaluated the scaffold under agitation by air in an MSUB from filtered air pumped into the cultivation chamber [42]. The scaffolds remained intact, showcasing their robustness and suitability for bioprocessing applications (See supplementary Video1).…”

“…The macrofluidic single use bioreactor fabricated as described in [42,44]. The flexible tube connected to the pump was introduced into the MSUB under the liquid surface, the flexible tube allowing air out of the MSUB is above the liquid phase.…”

“…Next, by changing the media composition, we have turned the bMSC-loaded-AVPC scaffold into 'lipid chunks'. Lastly, we show the potential utility of using this scaffold within a novel macrofluidic single-use bioreactor (MSUB) [42]. This approach promotes sustainability by utilizing renewable resources and is potentially scalable in a scale-out model, solving some of the technical challenges cultured meat faces [43].…”