1. Culture medium used during small interfering RNA (siRNA) transfection determines the maturation status of dendritic cells
- Author
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Essen, M.F. van, Schlagwein, N., Gijlswijk-Janssen, D.J. van, Anholts, J.D.H., Eikmans, M., Ruben, J.M., Kooten, C. van, and COMBAT Consortium
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0301 basic medicine ,Small interfering RNA ,Cell type ,T-Lymphocytes ,T cell ,Immunology ,Lymphocyte Activation ,Transfection ,Dendritic cells ,03 medical and health sciences ,RNA interference ,0302 clinical medicine ,medicine ,Humans ,Immunology and Allergy ,Gene silencing ,RNA, Small Interfering ,Cells, Cultured ,Chemistry ,Cell Differentiation ,Dendritic cell ,Culture Media ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Cell culture ,siRNA ,Cell maturation ,030215 immunology - 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.
- Published
- 2020
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