Photoelasticity-based evaluation of cellular contractile force for phenotypic discrimination of vascular smooth muscle cells
Photoelasticity-based evaluation of cellular contractile force for phenotypic discrimination of vascular smooth muscle cells
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DOI：
10.1038/s41598-019-40578-7
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发表时间：
2019-03
影响因子：
4.6
通讯作者：
Shukei Sugita;Eri Mizutani;Masatoshi Hozaki;Masanori Nakamura;Takeo Matsumoto
中科院分区：
文献类型：
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作者：
Shukei Sugita;Eri Mizutani;Masatoshi Hozaki;Masanori Nakamura;Takeo Matsumoto
血管平滑肌细胞（VSMC）有两种不同的表型：收缩型和合成型。这些表型之间的主要差异在于细胞产生的收缩力的大小。虽然牵引力显微镜（TFM）通常用于评估细胞收缩力，但这种方法需要复杂的预处理和足够柔顺的基底。为了评估的收缩力和表型的活VSMCs以最小的努力，并以独立的基板刚度的方式，我们提出了一种基于光弹性的方法，使用延迟，这是相关的第一和第二主应力和它们的方向之间的差异。结果表明，肌动蛋白丝与细胞中的高阻滞区域共定位，表明肌动蛋白丝促进了VSMC的阻滞。Calyculin A和Y-27632处理的细胞的阻滞倾向于更大和更小，分别比对照细胞。细胞牵引力与总细胞阻滞显著相关（r2= 0.38）。收缩型VSMCs（第2代）的延迟显著高于合成型VSMCs（第12代）。这些结果表明，细胞阻滞可用于评估细胞收缩力，从而确定VSMCs的表型。
Vascular smooth muscle cells (VSMCs) have two distinct phenotypes: contractile and synthetic. The major difference between these phenotypes lies in the magnitude of the contractile force produced by the cell. Although traction force microscopy (TFM) is often used to evaluate cellular contractile force, this method requires complex preprocessing and a sufficiently compliant substrate. To evaluate the contractile force and the phenotype of living VSMCs with minimal effort and in a manner independent of the substrate stiffness, we propose a photoelasticity-based method using retardation, which is related to the difference between the first and second principal stresses and their orientation. The results demonstrate that actin filaments co-localize with areas of high retardation in cells, indicating that the retardation of VSMCs is promoted by actin filaments. The retardation of cells treated with calyculin A and Y-27632 tended to be larger and smaller, respectively, than that of control cells. Cell traction force significantly correlates with total cell retardation (r2= 0.38). The retardation of contractile VSMCs (passage 2) was significantly higher than that of synthetic VSMCs (passage 12). These results indicate that cell retardation can be used to assess cell contractile force and, thus, determine the phenotype of VSMCs.