Department of Food Science, Aarhus University, Aarhus, Denmark.
Department of Food Science, Aarhus University, Aarhus, Denmark.
Food Res Int. 2023 Nov;173(Pt 1):113217. doi: 10.1016/j.foodres.2023.113217. Epub 2023 Jul 1.
Cultivated meat produced with primary muscle satellite cells (SCs) will need a continuous supply of isolated cell material from relevant animal donors. Factors such as age, sex, and breed, along with the sustainability and availability of donor animals, could determine the most appropriate donor type for an efficient production. In this study, we focus on the proliferation and differentiation of bovine SCs isolated from bull calf and dairy cow muscle samples. The proliferative performance of bull calf SCs was significantly better than SCs from dairy cows, however a dynamic differentiation assay revealed that the degree of fusion and formation of myotubes were similar between donor types. Furthermore, the proliferation of SCs from both donor types was enhanced using an in-house developed serum-free media compared to 10% FBS, which also delayed myogenic differentiation and increased final cell population density. Using gene chip transcriptomics, we identified several differentially expressed genes between the two donor types, which could help explain the observed cellular differences. This data also revealed a high biological variance between the three replicate animals within donor type, which seemed to be decreased when using our in-house serum-free media. With the use of the powerful imaging modalities of Cytation 5, we developed a novel high contrast brightfield-enabled label-free myotube quantification method along with a more efficient end-point fusion analysis using Phalloidin-staining. The results give new insights into the bovine SC biology and potential use of bull calves and dairy cows as relevant donor animals for cultivated beef cell sourcing. The newly developed differentiation assays will further enhance future research within the field of cultivated meat and SC biology.
利用原代肌肉卫星细胞（SCs）生产培养肉需要从相关动物供体中持续供应分离的细胞材料。供体动物的年龄、性别、品种、可持续性和可用性等因素可能会决定最适合高效生产的供体类型。在这项研究中，我们专注于从公牛犊和奶牛肌肉样本中分离的牛 SC 的增殖和分化。公牛犊 SC 的增殖性能明显优于奶牛 SC，然而，动态分化测定表明，供体类型之间的融合和肌管形成程度相似。此外，与 10% FBS 相比，使用内部开发的无血清培养基可增强两种供体类型的 SC 增殖，这也延迟了肌生成分化并增加了最终细胞群体密度。使用基因芯片转录组学，我们鉴定出两种供体类型之间的几个差异表达基因，这可以帮助解释观察到的细胞差异。该数据还揭示了供体类型内三个重复动物之间的高生物学变异性，当使用我们的无血清培养基时，这种变异性似乎会降低。使用 Cytation 5 的强大成像模式，我们开发了一种新的高对比度明场无标记肌管定量方法，以及使用鬼笔环肽染色进行更有效的终点融合分析。这些结果为牛 SC 生物学和作为培养牛肉细胞来源的公牛犊和奶牛的潜在用途提供了新的见解。新开发的分化测定将进一步增强培养肉和 SC 生物学领域的未来研究。
