Bioenergetics underlying single-cell migration on aligned nanofiber scaffolds
Bioenergetics underlying single-cell migration on aligned nanofiber scaffolds
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DOI：
10.1152/ajpcell.00221.2019
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发表时间：
2020-03-01
影响因子：
5.5
通讯作者：
Brown, David A.
中科院分区：
文献类型：
--
作者：
Padhi, Abinash;Thomson, Alexander H.;Brown, David A.
细胞迁移主要涉及无数的生理过程，包括形态发生、伤口愈合、组织修复和转移生长。迁移行为背后的生物能量学尚未完全理解，部分原因是细胞培养基的变化和实验细胞培养系统的利用不模拟生理连接的细胞外纤维网络。在这项研究中，我们评估了C2C12成肌细胞迁移的生物能量学和生产力的纤维连接蛋白涂覆的支架的控制直径和对齐，制造使用nonelectrospinning基于可调工程参数（STEP）平台。使用细胞呼吸、ATP合成、糖酵解或葡萄糖摄取的抑制剂确定各种代谢途径对细胞迁移的贡献。尽管对耗氧量有直接影响，但线粒体抑制仅适度降低细胞迁移速度，而糖酵解和细胞葡萄糖摄取抑制剂导致迁移显著减少。迁移代谢敏感性可根据细胞培养基中存在的底物进行修改。在半乳糖（而不是葡萄糖）中培养的细胞显示出对线粒体抑制的显著迁移敏感性。我们使用纳米网力显微镜来确定负责单细胞力生产的生物能量因素，并观察到无论是线粒体还是糖酵解抑制都不会改变单细胞力的生产。这些数据表明，成肌细胞迁移在很大程度上依赖于在传统培养基中生长的细胞中的糖酵解。这些研究对细胞迁移的原因、后果和假定的治疗方法有广泛的影响。
Cell migration is centrally involved in a myriad of physiological processes, including morphogenesis, wound healing, tissue repair, and metastatic growth. The bioenergetics that underlie migratory behavior are not fully understood, in part because of variations in cell culture media and utilization of experimental cell culture systems that do not model physiological connective extracellular fibrous networks. In this study, we evaluated the bioenergetics of C2C12 myoblast migration and force production on fibronectin-coated nanofiber scaffolds of controlled diameter and alignment, fabricated using a nonelectrospinning spinneret-based tunable engineered parameters (STEP) platform. The contribution of various metabolic pathways to cellular migration was determined using inhibitors of cellular respiration, ATP synthesis, glycolysis, or glucose uptake. Despite immediate effects on oxygen consumption, mitochondrial inhibition only modestly reduced cell migration velocity, whereas inhibitors of glycolysis and cellular glucose uptake led to striking decreases in migration. The migratory metabolic sensitivity was modifiable based on the substrates present in cell culture media. Cells cultured in galactose (instead of glucose) showed substantial migratory sensitivity to mitochondrial inhibition. We used nanonet force microscopy to determine the bioenergetic factors responsible for single-cell force production and observed that neither mitochondrial nor glycolytic inhibition altered single-cell force production. These data suggest that myoblast migration is heavily reliant on glycolysis in cells grown in conventional media. These studies have wide-ranging implications for the causes, consequences, and putative therapeutic treatments aimed at cellular migration.