Title: The impaction of nuclefection to dendritic cell
Abstract: Objective:To study the impaction of nuclefection to DC,including viability,expression of co-stimulatory molecules and the level of cytokines secretion by DC.Methods:p/IRES-EGFP was nuclefected into DC which was differentiated from PBMC in vitro.The un-nuclefected DC was used as control.typan blue exclusion assay was used to detect the viability of nuclefected DC.Flow cytometry was used to analyze the variation of CD80,CD83,CD86,HLA-DR and CD11c.The level of TNF-α and IL-12 produced by DC were measured by ELISA.Results:The viability was progressively decreased after nuclefection.EGFP could be detected in nuclefected-DC in 4 h after nulcefection,and the nuclefection efficiency was increasing gradually.The nuclefection efficiency was 56%±12% and 45±15% in 24 h and 48 h,respectively.The level of CD80,CD83 and CD86 decreased significantly in 48 h,but the level of HLA-DR and CD11c did not change.IL-12 and TNF-α produced by nuclefected DC were dramatically higher than that in control group(P0.05).Conclusion:Nuclefection is an effective method for DC transfection,and even can induce the secretion of proinflammatory cytokines.
Publication Year: 2011
Publication Date: 2011-01-01
Language: en
Type: article
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