PLEKHQ1基因敲除小鼠的永生化骨髓来源巨噬细胞系的建立  

Establishment of an immortalized bone marrow-derived macrophage cell line in PLEKHQ1 knockout mouse

作　　者：陆琤[1] 周晨辰[1] 张鹏飞[1] 张硌[1] 查玉华[1] 

机构地区：[1]军事医学科学院附属医院医学工程科,北京100071

出　　处：《中国医学装备》2017年第9期167-170,共4页China Medical Equipment

基　　金：国家自然科学基金(31400739)"PH结构域蛋白PLEKHQ1协调巨噬细胞迁移与激活的机制研究";军事医学创新基金(2015CXJJ29)"PLEKHQ1调控细菌性脓毒败血症的功能和机制研究"

摘　　要：目的:建立PLEKHQ1基因敲除小鼠的永生化骨髓来源巨噬细胞系,为PLEKHQ1基因功能的研究奠定基础。方法:用慢病毒感染的方法将包装质粒pCDH-SV40/GFP导入野生型和PLEKHQ1基因敲除小鼠的骨髓来源巨噬细胞,建立永生化细胞系;观察永生化细胞的生物学性状,用聚合酶链反应(PCR)检测目的基因的整合,用RT-PCR鉴定目的基因的表达,并对永生化和非永生化骨髓来源巨噬细胞的生长状况进行比较。结果:构建的骨髓来源巨噬细胞系已扩大培养并稳定传代,经鉴定,SV40LT抗原已整合入细胞并稳定表达。结论:通过慢病毒感染细胞的方法成功构建了PLEKHQ1基因敲除小鼠的永生化骨髓来源巨噬细胞系。Objective：To establish an immortalized bone marrow-derived macrophage（BMDM） cell line in PLEKHQ1 knockout mouse so as to pave the way for further studies on the function of PLEKHQ1 gene.Methods： Packaging plasmid pCDH-SV40/GFP was transfected into wild type mouse BMDM and PLEKHQ1 knockout mouse BMDM by the means of lentivirus infection so as to establish immortalized cell lines. The cell morphology of immortalized cell lines was observed and Polymerase chain reaction（PCR）was used to detect the integration of the target gene, and RT-PCR was used to identified the expression of target gene. The proliferation between the immortalized BMDM and non-immortalized BMDM was compared and analyzed in the finally.Results： The established immortalized BMDM cell line has achieved stable growth and serial propagation, and the results of identification showed that SV40LT antigen gene has been integrated in cell and been stably expressed.Conclusion： An immortalized BMDM cell line of PLEKHQ1 knockout mouse has been successfully established by the mean of. lentivirus infecting cells.

关 键 词：PLEKHQ1基因敲除小鼠 骨髓来源巨噬细胞 永生化 SV40LT抗原 

分 类 号：R516.8[医药卫生—内科学]