应用生物发光成像技术示踪植入的骨髓间充质干细胞  

In vivo Tracing of Transplanted Bone Marrow Mesenchymal Stem Cells with Bioluminescence Imaging

作　　者：边素艳[1] 盖鲁粤[2] 叶平[1] 杨月峰[1] 王华[3] 郭子宽[3] 王立生[3] 

机构地区：[1]解放军总医院老年心血管病二科 [2]解放军总医院心内科,北京100853 [3]军事医学科学院放射医学研究所实验血液学研究室,北京100850

出　　处：《中国实验血液学杂志》2009年第5期1307-1311,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助(编号30400182和30871018);国家863课题资助(编号2007AA02Z454)

摘　　要：以间充质干细胞(MSC)为基础的细胞治疗已进入临床试验阶段,用于多种组织修复,而移植细胞体内存活是影响疗效的重要因素。为了体内示踪MSC,构建了萤火虫荧光素酶基因(fluc)逆转录病毒表达载体pL(fluc)SN,并筛选出稳定表达fluc的GPE+86细胞克隆。将逆转录病毒上清转染C57小鼠来源的MSC,经G418筛选后得到稳定表达fluc的MSC。将2×106个标记好的细胞注射正常骨骼肌,应用生物发光技术检测移植后不同时间细胞的存活情况。结果表明,随着时间的延长,细胞存活数目逐渐减少;移植后1天细胞死亡率高达(43.2±11.7)%,3天后细胞存活(8.6±2.5)%,6天后仅为(5.4±3.1)%,10天时未检测到荧光信号。结论:生物发光成像技术可用于移植的MSC体内示踪,但是即使将MSC植入正常的骨骼肌组织,大部分注入的MSC也将在短期内死亡。Mesenchymal stem cell （MSC）-based cell therapy has shifted into clinical trials to repair the damage of various tissues. In this setting, the survival of the transplanted cells contributes critically to the therapeutic effectiveness. To investigate the in vivo tracing of MSCs, a recombinant retroviral vector carrying firefly-luciferase reporter gene [ pL （FLUC） SN ] was constructed and several GPE ~ 86 cell clones that stably expressed fluc were selected. The retroviral supernatants were collected and used to transfect MSC derived from C57 mice. The cells were then screened with G418 and the expression of the exogenous gene was identified by luciferase enzyme activity analysis. Labeled mouse MSCs （2 × 10^6） were injected into skeletal muscles, and the in situ expression was noninvasively tracked by in vivo bioluminescence imaging for 1, 3 and 6 days after transplantation. The results showed that the survival rates of the grafted cells dropped sharply with time, they were 57.2 ± 11.7 %, 8.6 ± 2.5 % and 5.4 ± 3.1% on day l, 3 and 6 after transplantation, and no fluorescent signals above background were detected on day 10. It is concluded that the method described above could be used for in vivo tracing of grafted cells. Furthermore, MSCs could not survive even transplanted into the none-ischemic skeletal muscles.

关 键 词：生物发光成像技术 骨髓间充质干细胞 细胞标记 细胞移植 

分 类 号：Q503[生物学—生物化学] Q811.5