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Mesenchymal Stem Cell Durotaxis Depends on Substrate Stiffness Gradient Strength

机译：间充质干细胞杜兰克斯取决于基材刚度梯度强度

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Mesenchymal stem cells (MSCs) respond to niche elasticity, which varies between and within tissues. Stiffness gradients result from pathological conditions but also occur through normal variation, e.g. muscle. MSCs undergo directed migration even in response to shallow stiffness gradients before differentiating. More refined gradients of both stiffness range and strength are needed to better understand mechanical regulation of migration in normal and disease pathologies. We describe polyacrylamide stiffness gradient fabrication using three distinct systems that generate stiffness gradients of physiological (1 Pa/µm), pathological (10 Pa/µm), and step (≥ 100Pa/um) strength spanning physiologically relevant stiffness for most soft tissue, i.e. 1–12 kPa. MSCs migrated to the stiffest region for each gradient. Time-lapse microscopy revealed that migration velocity scaled directly with gradient strength. Directed migration was reduced in the presence of the contractile agonist lysophosphatidic acid (LPA) and cytoskeletal-perturbing drugs nocodazole and cytochalasin; LPA- and nocodazole-treated cells remained spread and protrusive, while cytochalasin-treated cells did not. Untreated and nocodazole-treated cells spread in a similar manner, but nocodazole-treated cells had greatly diminished traction forces. These data suggest that actin is required for migration whereas microtubules are required for directed migration. The data also imply that in vivo, MSCs may have a more significant contribution to repairs in stiffer regions where they may preferentially accumulate.
机译：间充质干细胞（MSC）对生态位弹性有反应，生态位弹性在组织之间和组织内部变化。刚度梯度是由病理条件引起的，但也可能通过正常变化而发生，例如肌肉。甚至在分化之前，MSC响应于较浅的刚度梯度也会经历定向迁移。需要更精确的硬度范围和强度梯度，以更好地理解正常和疾病病理学中迁移的机械调节。我们描述了使用三种不同的系统生成聚丙烯酰胺刚度梯度的方法，这些系统会生成生理学（1 Pa / µm），病理学（10 Pa / µm）和跨步（≥100Pa / um）的刚度梯度，跨越大多数软组织的生理相关刚度，即1–12 kPa。对于每个梯度，MSC迁移到最硬的区域。延时显微镜显示，迁移速度与梯度强度成正比。在存在收缩激动剂溶血磷脂酸（LPA）和细胞骨架干扰药物诺考达唑和细胞松弛素的情况下，定向迁移减少。 LPA和诺考达唑处理的细胞保持扩散和突出，而细胞松弛素处理的细胞则没有。未经处理和经诺考达唑处理的细胞以相似的方式扩散，但经诺考达唑处理的细胞的牵引力大大降低。这些数据表明肌动蛋白是迁移所必需的，而微管是定向迁移所必需的。数据还暗示，在体内，MSC可能对它们优先聚集的较硬区域中的修复有更重要的贡献。

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