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Abstract Cultivated meat production requires bioprocess optimization to achieve cell densities that are multiple orders of magnitude higher compared to conventional cell culture techniques. These processes must maximize resource efficiency and cost-effectiveness by attaining high cell growth productivity per unit of medium. Microcarriers, or carriers, are compatible with large-scale bioreactor use, and offer a large surface-area-to-volume ratio for the adhesion and proliferation of anchorage-dependent animal cells. An ongoing challenge persists in the efficient retrieval of cells from the carriers, with conflicting reports on the effectiveness of trypsinization and the need for additional optimization measures such as carrier sieving. To surmount this issue, edible carriers have been proposed, offering the advantage of integration into the final food product while providing opportunities for texture, flavor, and nutritional incorporation. Recently, a proof of concept (POC) utilizing inactivated mycelium biomass derived from edible filamentous fungus demonstrated its potential as a support structure for myoblasts. However, this POC relied on a model mammalian cell line combination with a single mycelium species, limiting realistic applicability to cultivated meat production. This study aims to advance the POC. We found that the species of fungi composing the carriers impacts C2C12 myoblast cell attachment—with carriers derived fromAspergillus oryzaepromoting the best proliferation. C2C12 myoblasts effectively differentiated on mycelium carriers when induced in myogenic differentiation media. Mycelium carriers also supported proliferation and differentiation of bovine satellite cells. These findings demonstrate the potential of edible mycelium carrier technology to be readily adapted in product development within the cultivated meat industry.
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Uniaxially crumpled graphene as a platform for guided myotube formation
Abstract Graphene, owing to its inherent chemical inertness, biocompatibility, and mechanical flexibility, has great potential in guiding cell behaviors such as adhesion and differentiation. However, due to the two-dimensional (2D) nature of graphene, the microfabrication of graphene into micro/nanoscale patterns has been widely adopted for guiding cellular assembly. In this study, we report crumpled graphene, i.e., monolithically defined graphene with a nanoscale wavy surface texture, as a tissue engineering platform that can efficiently promote aligned C2C12 mouse myoblast cell differentiation. We imparted out-of-plane, nanoscale crumpled morphologies to flat graphene via compressive strain-induced deformation. When C2C12 mouse myoblast cells were seeded on the uniaxially crumpled graphene, not only were the alignment and elongation promoted at a single-cell level but also the differentiation and maturation of myotubes were enhanced compared to that on flat graphene. These results demonstrate the utility of the crumpled graphene platform for tissue engineering and regenerative medicine for skeletal muscle tissues.
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- PAR ID:
- 10154346
- Publisher / Repository:
- Nature Publishing Group
- Date Published:
- Journal Name:
- Microsystems & Nanoengineering
- Volume:
- 5
- Issue:
- 1
- ISSN:
- 2055-7434
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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