MSCs and meshes for Hernia - qPCR data.txt

Gene expression of cells infiltrated in the surgical meshes at 1 week and 1 month after surgical mesh implantation was analyzed by quantitative Real Time PCR (qPCR). qPCR was performed with commercial gene expression assays (Life Technologies, Thermo Fisher Scientific Inc.), using TaqMan Fast Advance Master Mix (Life Technologies, Thermo Fisher Scientific Inc.), in accordance with the protocol provided by the manufacturer. The amplification of cDNAs was performed in a QuantStudio 3 System (Applied Biosystems, Thermo Fisher Scientific) and the qPCR products were quantified by fluorescent method using the 2^-ΔCt expression. Duplicates of all samples were analyzed separately and normalized using HRPT1 as a reference gene.

本研究采用实时定量聚合酶链式反应（quantitative Real Time PCR，qPCR），对手术植入网片术后1周及1个月时，浸润于植入网片内的细胞的基因表达水平进行分析。实验使用Life Technologies（隶属于赛默飞世尔科技公司（Thermo Fisher Scientific Inc.））的商用基因表达检测试剂盒，搭配TaqMan Fast Advance预混液，严格遵循制造商提供的实验方案完成qPCR操作。cDNA的扩增在QuantStudio 3实时荧光定量PCR系统（Applied Biosystems，赛默飞世尔科技公司）中完成，并采用2^(-ΔCt)荧光定量法对qPCR产物进行定量分析。所有样本均设置重复样本独立检测，并以HRPT1作为内参基因完成数据归一化处理。