Establishment of CD200 stably-expressed U937 cell line / 中国免疫学杂志

ABSTRACT

Objective:To establish a human monocyte cell line U937 stably expressing CD200.Methods:The eukaryotic expression vector pcDNA3-CD200 containing human CD200 cDNA and vector pcDNA3 were transferred into human monocyte U937 cell line by electrotransfer.These two cell lines were selected by G418, and the selected cell lines were subcultured. U937 cell line was enclosed as control group.The expression of CD200 mRNA and protein was detected by RT-PCR and flow cytometry.Results:After G418 selection,U937 cell line in control group was died, and the cell lines transferring pcDNA3 and recombined pcDNA3-CD200 were subcultured over 30 generations.The CD200 mRNA expression in pcDNA3-CD200 group was confirmed with RT-PCR.The CD200 expression in U937-pcDNA3-CD200,U937-pcDNA3 and U937 were 77.20%,3.20% and 2.10%,compared with other two groups (F=133 996.40,P0.05).Conclusion:Our study provides foundation for mechanism of action for CD200 in future.The human monocyte series U937 cell line that expresses CD200 protein stably is established successfully by gene transfection method.