(B-72) High protein cell-based meat powder by high myoblast differentiation rate

Introduction:: Various alternative proteins are being developed to replace meat for solving some issues such as environmental, resource, animal welfare, and food shortage problems caused by traditional livestock [1]. However, alternative proteins (e.g., plant-based meat, microoraganisms, microalgae, and insects) have limitations such as nutritional value, safety, consumer perception, and organoleptic properties [2,3]. Hence, cell-based meat, also known as lab-grown or cultured meat, is emerging as a new protein source to overcome these limitations. The most important points in cell-based meat research are to reduce the cost for commercialization and to increase cell differentiation to resemble real meat [4-6]. Recently, many research papers in the field have proposed numorous forms of cultured meat in an effort to increase cell differentiation and reduce costs. Research on many types of cultured meat is being conducted, from using natural polymers or soy proteins as scaffolds to those made by stacking cell sheets [4,7,8]. Among various forms, we tried to increase commercial value by manufacturing cultured meat in the form of food powder that has excellent physicochemical stability and is easy to preserve and transport [9].

Materials and Methods:: The use of FBS in cultured meat production accounts for a large part of the high cost. FBS is a key material because it contains various substances necessary for cell culture, such as growth factors, hormones, and amino acids. However, it is expensive and has environmental and ethical concerns, so FBS should be reduced or replaced. In this study, we lowered the manufacturing cost and increased the differentiation efficiency by controlling the FBS concentration in the growth medium. The control group is G1: FBS 10% and experimental groups are G2: FBS 5%, G3: FBS 5% + C-PC (C-phycocyanin), a substance known to help myoblast proliferation [10].
The C2C12 (cell line of mouse myoblast) culture process includes proliferation and differentiation process. C2C12 were cultured with each (G1, G2, G3) growth medium for the first 7 days (proliferation), and cultured with the same DM for the next 5 days (differentiation). Then, the cells are collected and lyophilized to make a powder. As a result, we prepared a novel form of powdered cultured meat (called CPM; Cell-based meat powder) using highly differentiated cells with high commercialization potential. Finally, we analyzed the protein of CPM through BCA assay and proteome analysis, and proceeded with flavor analysis through GC-MS.

Results, Conclusions, and Discussions:: The proliferation of cells cultured in all three groups was similar, but the differentiation was significantly higher in the G2 and G3 groups (fusion index, G1: 21.9%, G2: 55.6%, G3: 58.8%). The amount of protein in the cells was G1 (0.73 μg/μl), G2 (1.0 μg/μl), and G3 (1.3 μg/μl), respectively, which was directly proportional to the fusion index. The cost-efficiency of the CPM was calculated by dividing the protein concentration by the price of the culture medium (Cost efficiency: G1: 1, G2: 1.76, G3:0.31). As a result, we selected G2 is the most suitable medium for CPM production because it can produce a large quantity of protein at the lowest price.
CPM is characterized by a high proportion of myotubes, resulting in high protein content (48.1%). This is higher than chicken breast (25.7%) and beef tenderloin (20.7%), which are well known as high-protein foods. Moreover, when analyzing the flavor of CPM and beef tenderloin using GC-MS, it was confirmed that a compound with a flavor similar to that of beef was detected, verifying the possibility of being used as meat.
Due to these characteristics, CPM is expected to offer exciting possibilities for the future of the cell-based meat industry. In this work, we only used muscle cells, but we plan to use cells such as fat, vascular cells, fibroblasts, and bone cells to enrich taste, nutrients, and flavor. In addition, since the range can be increased by utilizing cells of various animal species such as cattle, pigs, and chickens, CPM is expected to make a breakthrough in the cultured meat industry.

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