干细胞转染CMV-Luciferase-PGK-Puro慢病毒后化学发光成像  

Chemiluminescence imaging of conditions after tree shrews UC-MSCs are transfected with CMV-Luciferase-PGK-Puro lentivirus

作　　者：阮光萍[1] 刘菊芬[1] 李自安[1] 王金祥[1] 吕燕波[1] 庞荣清[1] 潘兴华[1] 

机构地区：[1]成都军区昆明总医院细胞生物治疗中心、干细胞与免疫细胞生物医药技术国家地方联合工程实验室、云南省细胞治疗技术转化医学重点实验室、云南省干细胞工程实验室、昆明市干细胞与再生医学研究重点实验室,昆明650032

出　　处：《西南国防医药》2016年第12期1378-1381,共4页Medical Journal of National Defending Forces in Southwest China

基　　金：国家科技支撑计划项目(2014BI01B0);国家973计划项目(2012CB5181060);云南省科技计划重点项目(2013CA005)

摘　　要：目的研究用化学发光成像的方法观察脐带间充质干细胞转染CMV-Luciferase-PGK-Puro慢病毒后的情况,代替活体成像仪的可行性。方法用CMV-Luciferase-PGK-Puro慢病毒转染树鼩脐带间充质干细胞,用最适浓度的嘌呤霉素筛选,存活的细胞用6孔板的3个孔培养,贴壁后,3个孔依次加入底物D-荧光素钾盐,用化学发光成像仪拍照,再用软件进行活体成像转换。将转染成功的细胞注入麻醉后的树鼩皮下,树鼩静脉注射底物。结果细胞加入底物后有生物发光,发光强度随底物作用时间延长而减弱。树鼩皮下也观察到发光细胞。结论 CMV-Luciferase-PGK-Puro慢病毒能成功转染树鼩脐带间充质干细胞,转染成功的细胞用于动物模型治疗后,可进行活体成像,观察细胞在动物体内的分布。objective To study the feasibility of chemiluminescence imaging instead of living imaging for observing the conditions after tree shrews umbilical cord mesenchymal stem cells （UC-MSCs） are transfected with CMV-Luciferase-PGK-Puro lentivirus. methods The tree shrew UC-MSCs transfected with CMV-Luciferase-PGK-Puro lentivirus were screened with puromycin of optimum concentration. The survival cells were cultured with three holes of six-hole plate. Upon adherence, the substrate d- luciferin potassium salt was successively added to the three holes. A picture was taken by chemiluminescence imager, after which living imaging conversion was conducted. The cells successfully transfected were injected under the skin of postanesthetic tree shrew, which were injected with substrate by intravenous injection, Results Upon the addition of substrate, the cells had bioluminescence; the luminousintensity weakened with the extension of substrate action time. Subcutaneous luminous cells were also observed under the skin of tree shrew. Collusion CMV-Lueiferase-PGK-Puro lentivirus may successfully transfeet tree shrews UC-MSCs. Successfully transfeeted cells used for the treatment of animal models can undergo living imaging to observe the distribution of cells in the animal body.

关 键 词：树鼩 脐带 间充质干细胞 CMV-Luciferase—PGK-Puro慢病毒 转染 化学发光成像 

分 类 号：R318.1[医药卫生—生物医学工程]