The Production of Fat-Containing Cultured Meat by Stacking Aligned Muscle Layers and Adipose Layers Formed From Gelatin-Soymilk Scaffold.
The Production of Fat-Containing Cultured Meat by Stacking Aligned Muscle Layers and Adipose Layers Formed From Gelatin-Soymilk Scaffold.
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DOI：
10.3389/fbioe.2022.875069
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发表时间：
2022
影响因子：
5.7
通讯作者：
Lin, Feng-Huei
中科院分区：
文献类型：
--
作者：
Li, Chi-Han;Yang, I-Hsuan;Ke, Cherng-Jyh;Chi, Chih-Ying;Matahum, Jefunnie;Kuan, Che-Yung;Celikkin, Nehar;Swieszkowski, Wojciech;Lin, Feng-Huei
组织工程培养肉已被提出作为一种新兴的肉类生产创新工艺，以克服畜牧业，气候变化和全球人口增长的严重后果。然而，目前培养肉缺乏有组织的组织结构，脂肪含量不足，生产成本高，这些都是当前面临的主要挑战。在这项研究中，使用明胶和豆奶合成了一种开发的支架，分别为C2 C12和3 T3-L1细胞中的肌生成和脂肪生成创造了一个友好的环境。含脂肪的培养肉通过将这些层交替堆叠而制成具有对齐的肌肉样层和脂肪样层。将表达肌球蛋白的肌肉样层和富含脂肪的脂肪样层夹在中间，形成含脂肪的肌肉组织。使用WST-1测定和活/死染色评价细胞毒性和细胞存活率。肌生成蛋白和肌球蛋白的表达证实了肌生成。在倒置显微镜、荧光显微镜和扫描电镜下观察肌管、肌原纤维和肌节。通过过氧化物酶体增殖物激活受体γ的蛋白表达来评估脂肪生成，并通过荧光显微镜和尼罗红染色来确定油滴积累。番红O染色检测细胞外基质分泌。在本研究中，通过添加前脂肪细胞制备具有肌肉样质地的培养肉，其中具有脂肪含量的多层肌肉样组织将产生多汁的培养肉。（A）制备明胶-豆奶溶液，然后置于PDMS模具上以制备具有平行微通道的GS支架。(B)将C2 C12成肌细胞和3 T3-L1前脂肪细胞接种在GS支架上以制备脂肪细胞覆盖的肌肉样层和脂肪样层。(C)然后将这两层一层一层地堆叠起来，组装成含有脂肪的培养肉。
Tissue engineered cultured meat has been proposed as an emerging innovative process for meat production to overcome the severe consequences of livestock farming, climate change, and an increasing global population. However, currently, cultured meat lacks organized tissue structure, possesses insufficient fat content, and incurs high production costs, which are the major ongoing challenges. In this study, a developed scaffold was synthesized using gelatin and soymilk to create a friendly environment for myogenesis and adipogenesis in C2C12 and 3T3-L1 cells, respectively. The fat containing cultured meat was fabricated with an aligned muscle-like layer and adipose-like layer by stacking these layers alternately. The muscle-like layer expressing myosin and the adipose-like layer abundant in fat were sandwiched to form fat containing muscle tissue. The cytotoxicity and cell survival rate were evaluated using the WST-1 assay and live/dead staining. Myogenesis was confirmed by the expression of myogenin and myosin. The myotubes, myofibrils, and sarcomeres were observed under an inverted microscope, fluorescence microscope, and scanning electron microscope. Adipogenesis was evaluated by protein expression of the peroxisome proliferator-activated receptor γ, and oil droplet accumulation was determined by fluorescence microscopy with Nile Red stain. Extracellular matrix secretion was examined by safranin-O staining. In this study, the cultured meat was prepared with muscle-like texture with the addition of pre-adipocyte, where the multilayered muscle-like tissues with fat content would produce juicy cultured meat. Scheme for cultured meat preparation (A) Gelatin–soymilk solution is prepared and then put on the PDMS mold to prepare a GS scaffold with parallel microchannels. (B) The C2C12 myoblasts and 3T3-L1 pre-adipocytes are seeded on the GS scaffold to prepare adipocyte-topped muscle-like layers and adipose-like layers. (C) The two layers are then stacked layer by layer to assemble into cultured meat containing fat.
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影响因子：
14
作者：
Nichol, Jason W.;Koshy, Sandeep T.;Bae, Hojae;Hwang, Chang M.;Yamanlar, Seda;Khademhosseini, Ali
通讯作者：
Khademhosseini, Ali
影响因子：
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Ironside, JW
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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