Gene silencing using small interfering ribonucleic acids (siRNA) is a powerful method to interfere with gene expression, allowing for the functional exploration of specific genes. siRNA interference can be applied in both cell lines, as well as in primary, non-dividing cell types like dendritic cells. However, the efficacy in different cell types is variable and requires optimization. Here, we showed that the type of culture medium used during lipid-based siRNA-mediated transfection acts as a critical factor, affecting dendritic cell activation. Transfection of immature monocyte-derived dendritic cells in RPMI medium, but not in IMDM, showed increased transcript levels of pro-inflammatory cytokines. Moreover, the expression of co-stimulatory molecules was enhanced, thereby increasing the T cell stimulatory capacity. Our data demonstrates that the choice of medium should be critically examined as one of the variables while optimizing cell transfection.

中文翻译：

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小干扰RNA（siRNA）转染过程中使用的培养基决定了树突状细胞的成熟状态。

使用小分子干扰核糖核酸（siRNA）进行基因沉默是一种强大的干扰基因表达的方法，可以对特定基因进行功能性探索。siRNA干扰可应用于两种细胞系，以及原代，非分裂型细胞（如树突状细胞）中。但是，在不同细胞类型中的功效是可变的，需要优化。在这里，我们表明在基于脂质的siRNA介导的转染过程中使用的培养基类型是影响树突状细胞激活的关键因素。未成熟单核细胞衍生的树突状细胞在RPMI培养基而不是IMDM中的转染显示促炎细胞因子的转录水平增加。此外，共刺激分子的表达增强，从而增加了T细胞的刺激能力。