【摘要】 目的探讨细胞培养基底刚度对心肌成纤维细胞增殖和粘附的影响。方法制备不同配比聚丙烯酰胺凝胶模拟不同基底刚度,采用细胞计数和MTT法检测细胞增殖,免疫荧光成像和免疫印迹方法检测细胞粘着斑分布及其相关蛋白表达。结果采用自重弯曲法测得0.03%、0.13%和0.26%聚丙烯酰胺凝胶的弹性模量为7～33Kpa。在此刚度范围内,随着基底刚度增加心脏成纤维细胞数量显著增加,粘着斑面积占细胞总面积的百分比增加,粘着斑纽蛋白的荧光强度增加,但纽蛋白和桩蛋白的表达量不变。结论基底刚度增加能促进新生大鼠心脏成纤维细胞的增殖和改变粘着斑蛋白的分布。更多还原

【Abstract】 Objective To observe the effects of substrate rigidity on proliferation and adhesion in neonatal rat cardiac fibroblasts.Methods Polyacrylamide gels with different ratio (Bis:Acr) were prepared to achieve different stiffness substrates for cell culture.The cell proliferation was determined by using cell counting and MTT assay.The distribution and expression of focal adhesion proteins were detected by immunofluorescence confocal image and Western Blotting.Results Young’s modulus of the substrate gel measured by the methods of micro-ball indentation,gravity stretching and special apparatus.Young’s modulus E value of the gel substrate ranged from 7 KPa to 33KPa.The variation of cell number,percentages of focal adhesion area in cell area,and fluorescence intensity of vinculin increases,but not expression of vinculin and paxillin,along with substrate stiffness increase.Conclusion Substrate stiffness increasing may contribute to cell proliferation and focal adhesion formation in neonatal rat cardiac fibroblasts.更多还原