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，资源下降的人口对负担得起的可持续食品和营养构成了重大挑战。一种资源有效的溶液是培养的肉，它是直接使用生物反应器直接培养动物细胞产生的真正动物肉。尽管基于脚手架的技术证明了制造切碎或非结构化肉类产品的可行性，但这些技术是有限的，无法产生厚实的结构化肉。这项探索性研究（急切）的早期概念赠款支持基础研究，旨在建立无脚手架的3D嵌入式生物打印技术，以使能够生产具有较厚和结构化特征的培养肉类。这项研究将探索在嵌入式肉类印刷过程中，在明胶复合材料的细胞基质浴中嵌入的肉类印刷过程中的成肌细胞的对齐和融合。结果将催化未来的培养肉类结构化切割量的扩大生产，并促进细胞农业，这是互补食品生产的未来，以实现可持续性，公共卫生和动物福利的利益。该项目还将刺激基于科学的生物印刷研究，以推动培养的磨牙制造，并通过生物印刷研究扩大了代表性不足的学生参与横切STEM领域的参与。这项研究的目的是了解挤出诱导的剪切力和构成肌肉纤维形成的挤压构成在构造构造的构造构成杂物层构造的影响。具体而言，成肌细胞将以嵌入式的方式印刷，对齐并伸展，以使成肌细胞融合为肌管，并在基于明胶复合材料的基于明胶复合的产量压力矩阵浴中进一步成熟。然后，将删除印刷的牺牲生物互联，形成灌注通道。虽然成肌细胞和脂肪细胞祖细胞的嵌入式3D打印将使印刷组织能够结构化，但由内皮和脂肪衍生的干细胞自组装的灌注通道和毛细血管会使印刷的组织变得浓密。从理论上讲，剪切力对印刷过程中成肌细胞对齐的影响将通过欧拉配方进行计算模型，而成肌细胞将宏观视为在肌细胞生物互中的线性弹性固体。建模结果将通过打印的成肌细胞的方向进行验证。接下来，将在定制的生物反应器中培养印刷肉类的组织期间研究环状张力诱导的对成肌细胞融合的作用，并通过通道灌注印刷的组织。由此产生的类似肉类的组织将以血管化，成肌细胞的分化以及肌管和肌纤维形成来表征。该奖项反映了NSF的法定任务，并被认为值得通过基金会的知识分子优点和更广泛的影响来通过评估来进行评估。

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