摘要
运用PiggyBac转座子系统,通过嘌呤霉素筛选构建稳定表达荧光素酶的U87MG细胞株(U87MG-FLUC)。通过流式细胞术检测U87MG-FLUC细胞株纯度达到100%。该细胞系传代后能稳定表达荧光素酶,且生物发光的强度与细胞个数成正比。将该稳定表达荧光素酶的细胞接种于免疫缺陷型小鼠的颅内,肿瘤接种后的7 d,利用活体成像仪对植瘤小鼠进行检测,确定肿瘤的大小以及位置。结果表明:接种了U87MG-FLUC细胞的裸鼠,均长出肿瘤;成功构建了表达荧光素酶的U87MG细胞株,且人脑胶质瘤细胞系的裸鼠肿瘤模型构建成功。
U87-MG cell line(U87MG-FLUC)stably expressing luciferase was constructed by puromycin selection using the PiggyBac transposon system.U87MG-FLUC cell line was 100%pure by flow cytometry;the cell line was able to stably express luciferase after passage,and the intensity of bioluminescence was directly proportional to the number of cells.The cells stably expressing luciferase were inoculated into the skull of immunodeficient mice.Seven days after tumor inoculation,tumor-implanted mice were detected using a live imager to determine the size and location of the tumor.Nude mice inoculated with U87MG-FLUC cells developed tumors.The results showed that the U87MG cell line expressing luciferase was successfully constructed,and a nude mouse tumor model of a human glioma cell line was successfully constructed.
作者
李雨莹
罗凯伦
邹宗幸
刘滨磊
汪洋
LI Yuying;LUO Kailun;ZOU Zongxing;LIU Binlei;WANG Yang(School of Biological Engin.and Food Science,Hubei Univ.,of Tech.Wuhan 430068,China)
出处
《湖北工业大学学报》
2020年第5期75-77,87,共4页
Journal of Hubei University of Technology
