Construction of folate-modified and CD47 siRNA-loaded exosomes
[Objective]To construct folate-modified and CD47 siRNA-loaded exosomes.[Method]NIH/3T3 cells were cultured in the medium with different concentrations of folate,and the folate content was detected with immunostaining assay.The cell viability was detected by CCK-8 assay.The folate-modified exosomes were isolated by step-by-step centrifuga-tion.CD47 siRNA was loaded into the exosomes by electrotransfection.The loading efficiency was measured by flow cytometry.The knockdown effect of CD47 was evaluated by RT-qPCR.[Result]When the concentration of folate was lower than 20 µg/mL,the cell viability of NIH/3T3 was not influenced significantly and the cell membrane was rich in folate.The diameter of ex-osomes was about 125 nm,and rich in CD63 and CD9 protein expression.The electrotransfection efficiency of CD47 siRNA into exosomes was about 86.9％,and this drug-loading exosome knocked down the mRNA expression of CD47 to 18％.[Conclu-sion]CD47 siRNA loaded and folate-modified exosomes can knockdown the protein expression of CD47 of cancer cells.
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