Nofima - Norwegian Institute of Food, Fisheries and Aquaculture Research, P.O. Box 210, N-1431, Ås, Norway.
Food Funct. 2020 Mar 26;11(3):2477-2488. doi: 10.1039/c9fo02690h.
The most significant cost driver for efficient bio-production of edible animal proteins is the cell culture media, where growth factors account for up to 96% of the total cost. The culture media must be serum-free, affordable, contain only food-grade ingredients, be efficient to promote cell growth and available in massive quantities. The commercially available serum substitutes are expensive and not necessarily food-grade. Identifying inexpensive food-safe alternatives to serum is crucial. By-products from food production are available in massive quantities, contain potential factors that can promote growth and are promising ingredients for serum replacement. The main goal of this study was to explore if food-grade by-product materials can be used as growth promoting agents in skeletal muscle cell culture to develop a tailor-made serum free media. Different by-products, including chicken carcass, cod backbone, eggshell membrane, egg white powder and pork plasma were enzymatically or chemically hydrolyzed. The hydrolysates in addition to lyophilized pork plasma and yeast extract were further characterized by size-exclusion chromatography, elemental combustion analysis and degree of hydrolysis. The materials were used as supplement to or replacement of commercial serum and further evaluated for their effect on metabolic activity, cell proliferation and cell cytotoxicity in muscle cells cultured in vitro. Our results indicate that none of the materials were cytotoxic to the skeletal muscle cells. Hydrolysates rich in peptides with approximately 2-15 amino acids in length were shown to improve cell growth and metabolic activity. Of all the materials tested pork plasma hydrolysates and yeast extract were the most promising. Pork plasma hydrolysates increased metabolic activity by 110% and cell proliferation with 48% when cultured in serum-free conditions for 3 days compared with control cells cultured with full serum conditions. Most interestingly, this response was dependent on both material and choice of enzyme used. We suggest that these materials have the potential to replace serum during cultivation and as such be included in a tailor-made serum-free media.
高效生产可食用动物蛋白的生物过程中，最主要的成本驱动因素是细胞培养基，其中生长因子占总成本的 96%。培养基必须不含血清、价格合理、只包含食品级成分、高效促进细胞生长且能大量供应。市售的血清替代品不仅昂贵，而且不一定是食品级的。寻找便宜且安全的血清替代品至关重要。食品生产的副产物大量存在，其中可能含有促进生长的因子，是替代血清的有前途的成分。本研究的主要目的是探索食品级副产物材料是否可以用作骨骼肌细胞培养中的生长促进剂，以开发定制的无血清培养基。不同的副产物，包括鸡骨架、鳕鱼骨、蛋壳膜、蛋白粉和猪血浆，经酶解或化学水解。水解产物以及冻干猪血浆和酵母提取物进一步通过凝胶过滤色谱、元素燃烧分析和水解度进行了表征。将这些材料作为补充或替代商业血清使用，并进一步评估它们对体外培养的肌肉细胞代谢活性、细胞增殖和细胞毒性的影响。我们的结果表明，这些材料对骨骼肌细胞均无细胞毒性。富含 2-15 个氨基酸的短肽水解产物可改善细胞生长和代谢活性。在所有测试的材料中，猪血浆水解产物和酵母提取物最有前景。与完全用血清培养的对照细胞相比，在无血清条件下培养 3 天时，猪血浆水解产物可使代谢活性提高 110%，细胞增殖提高 48%。最有趣的是，这种反应依赖于所用材料和酶的选择。我们认为这些材料有可能在培养过程中替代血清，从而被纳入定制的无血清培养基中。
