电穿孔对HT29细胞系与微图案细胞外基质蛋白(纤连蛋白)相互作用影响的研究

H. Mamman, M. Jamil, M. N. Adon
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引用次数: 1

摘要

微接触印刷(MCP)是一种允许基质或表面与细胞外基质(ECM)蛋白以明确定义的方式自由功能化的方案。MCP可用于调节细胞在基质上的粘附几何形状和控制伤口愈合过程。在这项研究中,人结肠癌细胞系HT29在纤维连接蛋白微接触印刷图案上生长,重复光栅宽度为25µm, 50µm和100µm,生长48小时。然后计算细胞与图案底物的对齐,其中0°表示与图案100%对齐。这样做的目的是找到那些模式,刺激细胞排列的最佳程度。定量分析表明,HT29细胞最容易向50µm和100µm方向排列,平均排列角度分别为4.8°±1.5SD和5.7°±1.4SD。另一方面,HT29在25um和未涂布纤维连接蛋白基板上的平均对准角分别为33.4°±8.4SD和51.1°±9.5SD,对准角较差。此外,采用50µm的印花图案来研究电场对HT29与纤维连接蛋白图案衬底对齐的影响。电穿孔和非电穿孔HT29细胞在50µm图案化底物上培养48小时。结果表明,电场处理的细胞的平均角度为4.2°±1.4SD,未处理的细胞的平均角度为4.8°±1.5SD,两者之间的对准度无显著差异。然而,与未处理的细胞在44小时后排列相比,经电处理的细胞排列更快(在播种后31小时内)。因此,研究结果表明,微接触印刷技术结合脉冲电场可以提供一种潜在的更快的方法来控制伤口愈合过程。
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Studying the influence of electroporation on HT29 cell line interaction with micro-patterned extracellular matrix protein (fibronectin)
Micro-contact printing (MCP) is a scheme that allows a substrate or surface to be functionalized freely with a protein of extracellular matrix (ECM), in a well-defined manner. MCP can be used to regulate cell adhesion geometry on a substrate and in controlling wound healing process. In this study, human colon cancer cell line, HT29 were grown on a micro-contact printed pattern of fibronectin with repeat gratings of 25µm, 50µm, and 100µm wide, for 48 hours. The cells alignments to the patterned substrates were then computed, where 0° means 100% alignment to the pattern. This was done with the purpose of finding those pattern that stimulated the best degree of cell alignment. The quantitative analysis of the study revealed that HT29 cells aligned most readily to the 50µm and 100µm pattern with a mean angle of alignment of 4.8°±1.5SD and 5.7°±1.4SD respectively. On the other hand, HT29 poorly aligned to 25um and the un-patterned fibronectin-coated substrate with a mean angle of alignment of 33.4°±8.4SD and 51.1°±9.5SD respectively. Furthermore, the 50µm stamp pattern was used to investigate the influence of electric field on the HT29 alignment to the fibronectin patterned substrate. Electroporated and non-electroporated HT29 cells were cultured on the 50µm patterned substrates for 48 hours. The result revealed that there is no significant difference in the degree of alignment between the electrically treated with a mean angle of 4.2°±1.4SD and the untreated cells with a mean angle of 4.8°±1.5SD. However electrically treated cell aligned faster (within 31 hours of seeding) as compared to the untreated cells that aligned after 44 hours of seeding. Hence, the result of the study revealed that Micro-contact printing technique together with pulse electric field could offer a potential faster method of controlling wound healing process.
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