Construction of a SW620 Cell line Stably Expressing Human CD 19 and Fluorescent Protein based on Retroviral Packaging Technology
Objective:To construct SW620 cell line expressing human CD 19,enhanced Green fluorescent protein(EGFP)and lu-ciferase(LUC).Methods:Construct the MFG-CD19 plasmid,package it with a retroviral vector,and co transfect it with the MFG-EGFP-P2A-LUC retroviral vector to SW620 cells.Select a stable and strongly expressed monoclonal cell line SW620-CD19-EGFP-LUC and expand the culture.The expression of CD 19 on the cell was detected by flow cytometry,and the expres-sion of luciferase on the cell line surface and cell line function were identified by luciferase assay.Results:The percentage of CD19-posi-tive cells and EGFP-positive cells was 99.9％and 99.2％,respectively,and the green fluorescence could be observed in the Fluorescence microscope The results of qPCR showed that the expression level of CD 19 in the cell lines was significantly up-regulated compared with the original cell lines.Conclusions:The SW620 cell line that can stably express human CD 19 and fluorescent protein was successfully constructed through retroviral vector packaging technology.
HumanCD19SW620 cellRetroviral vectorColorectal cancer
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