The Construction of CD64(FcγRI)Coated Nanovesicle From the Cytoplast of Human Umbilical Cord Mesenchymal Stem Cells(HUC-MSCs)
Objective:The nanovesicle derived from hum、an umbilical cord mesenchymal stem cells(HUC-MSCs)can be used for drug targeting through a variety of methods.The aim of this study is to construct the CD64(FcγRI)coated nanovesicle from the HUC-MSCs,and to detect the efficiency of antibodies(IgG1/IgG3)loading and cell entry of the nanovesicle.Methods:HUC-MSCs were isolated by tissue-cutting and cell culture method;A pLV-CD64 lentivirus packaging vector was established and packaged in the lentivirus;The lentivirus was transfected into HUC-MSCs,and the cell line that stably express CD64 was selected;The CD64 expressing HUC-MSCs was denuclearizated using the cytochalasin B co-incubation/centrifugation method;The nanovesicle with a diameter of ap-proximately 200 nm was generated by ultrasound and gradient extrusion methods.The nanovesicle was co-incubated with anti-EGFR IgG1 antibodies for antibody loading,and the entry efficiency of the nanovesicle was investigated in A549 cells.Results:The nanovesicle with the diameter of 195±82 nm,was obtained using the cytoplast of HUC-MSCs.The nanovesicle was stably expressing CD64 and successfully loaded with EGFR antibody;The modification effectively improved the entry efficiency of the nanovesicle for target cells.Conclusion:A nanovesicle with the capable of loading IgG1 antibodies was constructed,which could efficiently improve its entry effi-ciency for target cells.
Human umbilical cord mesenchymal stem cells(HUC-MSCs)CytoplastNanovesicleCD64(FcγRI)
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