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. 2020 Apr:479:112748.
doi: 10.1016/j.jim.2020.112748. Epub 2020 Jan 17.

Culture medium used during small interfering RNA (siRNA) transfection determines the maturation status of dendritic cells

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Free article

Culture medium used during small interfering RNA (siRNA) transfection determines the maturation status of dendritic cells

Mieke F van Essen et al. J Immunol Methods. 2020 Apr.
Free article

Abstract

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.

Keywords: Cell maturation; Dendritic cells; RNA interference; Transfection; siRNA.

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