EAGER: 3D Printing of Aligned Muscle Fibers for Thick Structured Meat Production

EAGER：用于厚结构肉生产的对齐肌肉纤维的 3D 打印

• 批准号: 2233814
• 负责人: Yong Huang
• 金额: $ 30万
• 依托单位: University of Florida
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2233814&HistoricalAwards=false
• 关键词: EAGER; 3D; Printing; Aligned; Muscle

The continued growth of human populations with declined resources has imposed a significant challenge to affordable and sustainable foods and nutrition. One resource-efficient solution is cultured meat, which is genuine animal meat produced by cultivating animal cells directly using a bioreactor. Though scaffold-based technologies have demonstrated the feasibility of making minced or unstructured meat products, such technologies are limited and cannot produce thick structured meat. This EArly-concept Grant for Exploratory Research (EAGER) supports fundamental research that aims to establish a scaffold-free 3D embedded bioprinting technology to enable the production of cultured meat with centimeter thick and structured features. The study will explore the alignment and fusion of myoblasts during embedded meat printing in a gelatin composite-based cellular matrix bath. The results will catalyze future scale-up production of thick structured cuts of cultured meat and promote cellular agriculture as the future of complementary food production for the benefits of sustainability, public health, and animal welfare. The project will also stimulate science-based bioprinting research to advance cultured-meat manufacture and broaden the participation of underrepresented students in crosscutting STEM fields via the bioprinting study.The objective of this research is to understand the effects of extrusion-induced shear force and post-printing tension on the formation of aligned muscle fibers from myoblasts during embedded printing of thick multicellular structured meat-like tissues. Specifically, myoblasts will be printed in an embedded manner, aligned, and stretched for myoblast fusion to be myotubes and further matured as myofibers in the gelatin composite-based yield-stress matrix bath. The printed sacrificial bioink will then be removed to form perfusable channels. While embedded 3D printing of myoblasts and adipocyte progenitor cells will enable printed tissues to be structured, the perfusable channels and capillaries self-assembled by endothelial and adipose-derived stem cells will enable the printed tissues to grow thick. Theoretically, the effect of shear force on the myoblast alignment during printing will be computationally modeled using the Eulerian formulation and myoblasts will be macroscopically treated as a linear elastic solid in the myoblast bioink. The modeling results will be validated with the orientation of the printed myoblasts. Next, the cyclic tension-induced effect on myoblast fusion will be investigated during the culturing of the printed meat-like tissues in a customized bioreactor, and the printed tissues will be perfused via the channels. The resulting meat-like tissues will be characterized in terms of vascularization, myoblast differentiation as well as myotube and myofiber formation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

人口持续增长而资源减少，对负担得起且可持续的食品和营养提出了重大挑战。一种资源高效的解决方案是培养肉，它是通过直接使用生物反应器培养动物细胞而产生的真正的动物肉。尽管基于支架的技术已经证明了制造碎肉或非结构化肉产品的可行性，但此类技术是有限的并且不能生产厚的结构化肉。这项早期概念探索性研究资助 (EAGER) 支持基础研究，旨在建立无支架 3D 嵌入式生物打印技术，以生产具有厘米厚和结构化特征的培养肉。该研究将探索在基于明胶复合材料的细胞基质浴中嵌入肉类打印过程中成肌细胞的排列和融合。研究结果将促进未来厚结构培养肉的大规模生产，并促进细胞农业成为补充食品生产的未来，以造福可持续发展、公共健康和动物福利。该项目还将刺激基于科学的生物打印研究，以推进培养肉生产，并通过生物打印研究扩大代表性不足的学生对交叉 STEM 领域的参与。这项研究的目的是了解挤压引起的剪切力和后力的影响-在厚的多细胞结构肉样组织的嵌入打印过程中，打印张力对成肌细胞形成排列的肌肉纤维的影响。具体来说，成肌细胞将以嵌入的方式打印、排列和拉伸，以使成肌细胞融合成肌管，并在基于明胶复合材料的屈服应力基质浴中进一步成熟为肌纤维。然后，打印的牺牲生物墨水将被去除以形成可灌注通道。虽然成肌细胞和脂肪祖细胞的嵌入式 3D 打印将使打印的组织结构化，但由内皮和脂肪干细胞自组装的可灌注通道和毛细血管将使打印的组织变厚。理论上，打印过程中剪切力对成肌细胞排列的影响将使用欧拉公式进行计算建模，并且成肌细胞将在宏观上被视为成肌细胞生物墨水中的线弹性固体。建模结果将通过打印的成肌细胞的方向进行验证。接下来，将研究在定制的生物反应器中培养打印的肉样组织期间循环张力诱导的对成肌细胞融合的影响，并且打印的组织将通过通道进行灌注。由此产生的肉样组织将在血管化、成肌细胞分化以及肌管和肌纤维形成方面进行表征。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Laponite nanoclay-modified sacrificial composite ink for perfusable channel creation via embedded 3D printing

• DOI: 10.1016/j.compositesb.2023.110851
• 发表时间: 2023-08
• 期刊: Composites Part B: Engineering
• 作者: B. Ren;Kaidong Song;Yunxia Chen;W. Murfee;Yong Huang