鉴定慢病毒携带IL-21转导人脐带间充质干细胞及IL-21活性初探  

Identification of human umbilical card mesenchymal stem cells infected with lentivirus vector encoding IL-21 and preliminary investigation into the functional activity of expressed IL-21

作　　者：张云霞[1] 王净[1] 任慕兰[1] 陈登宇[1] 陈峻崧[2] 王小英[2] 李淼[2] 窦骏[2] 

机构地区：[1]东南大学附属中大医院妇产科 [2]东南大学医学院病原生物学与免疫学系,南京210009

出　　处：《现代免疫学》2014年第5期358-362,共5页Current Immunology

基　　金：国家自然科学基金(81071769;81202372)

摘　　要：建立慢病毒携带IL-21转导人脐带间充质干细胞方法,并鉴定IL-21活性。以pRSC-IL-21质粒为模板扩增IL-21基因,插入到pHAGE-CMV-MCS-IzsGreen中,构建成慢病毒载体pHAGE-IL-21,将其包装为成熟慢病毒感染HEK 293T细胞。Western blot分析重组慢病毒感染hUCMSC中IL-21的表达,FCM法检测培养上清对脾细胞的增殖作用。结果显示正确构建了携带IL-21基因的慢病毒表达载体,包装的慢病毒感染HEK 293T细胞后可见绿色荧光表达。含IL-21慢病毒感染的hUCMSC中可检测到IL-21表达,分泌在培养上清中的IL-21具有促使脾细胞增殖活性。To construct the human umbilical card mesenehymal stem cells（hUCMSCs）infected with the recombinant lentivirus containing IL-21 gene,and to provide a new method for targeted treatment of cancer,the fragment of IL-21 gene from vector pRSC-IL-21 was cloned into the lentivirus vector pHAGE-CMV-MCS-IzsGreen and the lentivirus was packaged and subsequently transfected into the human embryo kidney HEK 293 T cells.The expression of IL-21 was detected in the recombinant lentivirus infected hUCMSCs by Western blot analysis.The results showed that the lentivirus vector carrying the IL-21 gene was established successfully.The expression of green fluorescent protein was observed under a fluorescence microscope after the recombinant lentivirus was cotransfected into HEK 293 T cells.The expression of IL-21 was detected from the recombinant lentivirus infected hUCMSCs and the expressed IL-21 in the supernatant could induce spleen cell proliferation.These results indicate that the correctly constructed recombinant lentivirus containing IL-21 gene could infect the hUCMSCs that express functional IL-21.

关 键 词：人脐带间充质干细胞 IL-21 慢病毒 靶向治疗 

分 类 号：R722.12[医药卫生—儿科]