Combe Michelle, Isaac Kathy Sharon, Potter Greg, Sokolenko Stanislav
Process Engineering and Applied Science, Dalhousie University, 5273 DaCosta Row, PO Box 15000, Halifax, B3H 4R2, NS, Canada.
Lipiferm Scientific, Halifax, NS, Canada.
Curr Res Food Sci. 2024 Sep 21;9:100855. doi: 10.1016/j.crfs.2024.100855. eCollection 2024.
Cultivated meat products, generated by growing isolated skeletal muscle and fat tissue, offer the promise of a more sustainable and ethical alternative to traditional meat production. However, with cell culture media used to grow the cells accounting for 55-95% of the overall production cost, achieving true sustainability requires significant media optimization. One means of dealing with these high costs is the use of low-cost complex additives such as hydrolysates to provide a wide range of nutrients, from small molecules (metabolites) to growth factors and peptides. Despite their potential, most hydrolysate products remain poorly characterized and many are thought to suffer from persistent issues of high batch-to-batch variability. Although there have been a number of isolated efforts to determine metabolic profiles for a handful of hydrolysate products, we present the first attempt at a more comprehensive metabolomic characterization of nine different products (four plant and five yeast-based) from two to four different lots each. NMR analysis identified 90 unique metabolites, with only 15 metabolites common to all hydrolysate products (including eight of the nine essential amino acids), and 16 metabolites found in only a single hydrolysate product. The different hydrolysate products were found to have substantial differences in metabolite concentrations (as a fraction of overall mass), ranging from a high of 43% in yeast extract to a low of 14% in soy hydrolysates. The proportion of various metabolites also varied between products, with carbohydrate concentrations particularly high in soy hydrolysates and nucleosides more prominent in two of the yeast products. Overall, yeast extract generally had higher metabolite concentrations than all the other products, whereas both yeast extract and cotton had the largest variety of metabolites. A direct calculation of batch-to-batch variability revealed although there are significant differences between lots, these are largely driven by a relatively small fraction of compounds. This report will hopefully serve as a useful starting point for a more nuanced consideration of hydrolysate products in cell culture media optimization, both in the context of cultivated meat and beyond.
通过培养分离的骨骼肌和脂肪组织生产的人造肉产品，有望成为一种比传统肉类生产更具可持续性和道德性的选择。然而，用于培养细胞的细胞培养基占总生产成本的55%-95%，要实现真正的可持续性，需要对培养基进行重大优化。应对这些高成本的一种方法是使用低成本的复合添加剂，如水解产物，以提供从小分子（代谢物）到生长因子和肽等广泛的营养物质。尽管它们具有潜力，但大多数水解产物的特性仍不清楚，许多产品被认为存在批次间差异大的持续问题。虽然已经有一些单独的努力来确定少数水解产物的代谢谱，但我们首次尝试对来自两到四个不同批次的九种不同产品（四种植物基和五种酵母基）进行更全面的代谢组学表征。核磁共振分析确定了90种独特的代谢物，所有水解产物中只有15种代谢物是共有的（包括九种必需氨基酸中的八种），还有16种代谢物仅在一种水解产物中发现。不同的水解产物在代谢物浓度（占总质量的比例）上存在显著差异，从酵母提取物中的43%到大豆水解产物中的14%不等。各种代谢物的比例在不同产品之间也有所不同，大豆水解产物中的碳水化合物浓度特别高，而两种酵母产品中的核苷更为突出。总体而言，酵母提取物的代谢物浓度通常高于所有其他产品，而酵母提取物和棉花的代谢物种类最多。对批次间差异的直接计算表明，虽然批次之间存在显著差异，但这些差异在很大程度上是由相对较少的一部分化合物驱动的。本报告有望成为在人造肉及其他领域的细胞培养基优化中更细致地考虑水解产物的有用起点。
