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Cyclic Stretch Effects on Adipose-Derived Stem Cell Stiffness Morphology and Smooth Muscle Cell Gene Expression

机译:循环拉伸对脂肪来源干细胞的刚度形态和平滑肌细胞基因表达的影响

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摘要

Recent investigations consider adipose-derived stem cells (ASCs) as a promising source of stem cells for clinical therapies. To obtain functional cells with enhanced cytoskeleton and aligned structure, mechanical stimuli are utilized during differentiation of stem cells to the target cells. Since function of muscle cells is associated with cytoskeleton, enhanced structure is especially essential for these cells when employed in tissue engineering. In this study by utilizing a custom-made device, effects of uniaxial tension (1Hz, 10% stretch) on cytoskeleton, cell alignment, cell elastic properties, and expression of smooth muscle cell (SMC) genes in ASCs are investigated. Due to proper availability of ASCs, results can be employed in cardiovascular engineering when production of functional SMCs in arterial reconstruction is required. Results demonstrated that cells were oriented after 24 hours of cyclic stretch with aligned pseudo-podia. Staining of actin filaments confirmed enhanced polymerization and alignment of stress fibers. Such phenomenon resulted in stiffening of cell body which was quantified by atomic force microscopy (AFM). Expression of SM α-actin and SM22 α-actin as SMC associated genes were increased after cyclic stretch while GAPDH was considered as internal control gene. Finally, it was concluded that application of cyclic stretch on ASCs assists differentiation to SMC and enhances functionality of cells.
机译:最近的研究认为,脂肪干细胞(ASCs)是临床治疗中有希望的干细胞来源。为了获得具有增强的细胞骨架和对齐结构的功能性细胞,在干细胞分化为靶细胞的过程中利用了机械刺激。由于肌肉细胞的功能与细胞骨架有关,因此在组织工程中使用增强的结构对于这些细胞尤为重要。在这项研究中,利用定制设备,研究了单轴张力(1Hz,10%拉伸)对ASC中细胞骨架,细胞排列,细胞弹性和平滑肌细胞(SMC)基因表达的影响。由于ASC的适当可用性,当需要在动脉重建中生产功能性SMC时,可将结果用于心血管工程。结果表明,在经过24小时的循环拉伸后,细胞具有定向的伪足。肌动蛋白丝的染色证实增强的聚合和应力纤维的排列。这种现象导致细胞体变硬,这通过原子力显微镜(AFM)定量。循环拉伸后,SMα-肌动蛋白和SM22α-肌动蛋白作为SMC相关基因的表达增加,而GAPDH被认为是内部控制基因。最后,得出结论,在ASC上应用循环拉伸有助于向SMC分化并增强细胞功能。

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