Speaker
Description
Introduction
The development of sustainable, structured cultivated meat represents a major challenge where tissue engineering and food technology must converge. Despite important achievements in recent years — including the isolation and expansion of satellite cells, and the blending of animal cells with plant-based matrices — replicating the hierarchical microarchitecture of real meat remains a critical bottleneck. Here, we present our advances toward the biofabrication of microarchitected meat using chaotic bioprinting strategies.
Materials and methods
We employed multimaterial chaotic bioprinting strategies to fabricate gelatin-based, meat-like constructs. Custom printheads equipped with 2 to 4 inlets and fitted with 2 to 6 Kenics static mixing (KSM) elements were used to induce chaotic flows and generate highly ordered microarchitectures within gelatin-based hydrogel filaments. Murine myoblasts (C2C12) and fibroblasts (3T3) were used as model cell types.
The progression of the meat-like constructs, from initial printing through to tissue maturation, was evaluated through a series of assays. Cell viability was assessed using live/dead fluorescence staining, while metabolic activity was quantified via PrestoBlue assays and glucose uptake measurements. Muscle-specific gene expression was analyzed to monitor tissue differentiation, and immunostaining was performed to assess protein-level expression of relevant muscle maturation biomarkers.
Results and discussion
We demonstrate that simple chaotic printing, utilizing static mixer-equipped printheads, enables the extrusion of hydrogel filaments loaded with murine myocytes (C2C12 cells) and containing internal void channels1. These channels facilitate myocyte alignment, promote multinucleation, enhance nutrient diffusion to the filament core, and support the formation of highly organized muscle-like fibers.
Through multimaterial chaotic bioprinting2, we also introduce reinforcing scaffolding inks during the extrusion process. These scaffolds enhance the mechanical stability of the printed constructs, enabling long-term culture under dynamic perfusion conditions without compromising structural integrity.
Additionally, the integration of fibroblast-laden inks during chaotic coextrusion adds a biologically active connective tissue-like component to the engineered constructs. Our results show that fibroblast coculture accelerates the consolidation and maturation of muscle-like fibers, supporting tissue organization and contributing to extracellular matrix deposition.
Finally, we describe variations in chaotic bioprinting materials and strategies aimed at improving scalability and manufacturability, setting the groundwork for future translation of microarchitected cultivated meat production.
Conclusion
Our results demonstrate that chaotic bioprinting is a promising platform to engineer complex, organized, and scalable cell-based meat tissues, bridging biological fidelity with manufacturing practicality. We envision the translatability of these techniques to bovine cells to fabricate micro-architected “sashimi” size pieces of eatable meat-like materials in the near future.
References:
Bolívar-Monsalve EJ et al. One-Step Bioprinting of Multi-Channel Hydrogel Filaments Using Chaotic Advection: Fabrication of Pre-Vascularized Muscle-Like Tissues. 2022. Adv Healthc Mater 11, (24), 2200448.
Ceballos-González CF et al. 2023. Plug-and-Play Multimaterial Chaotic Printing/Bioprinting to Produce Radial and Axial Micropatterns in Hydrogel Filaments. Adv Mater Technol 8, (17), 2202208.
Disclosure:
Grissel Trujillo de Santiago and Mario Moisés Alvarez have submitted patent applications protecting different aspects of a printing/bioprinting technique based on the use of chaotic flows.
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