不同输注途径对荧光素酶基因标记骨髓间充质干细胞体内分布的影响

发布时间：2018-01-20 08:48

本文关键词：组织工程干细胞股骨骨髓干细胞骨髓间充质干细胞股骨注射生物发光红荧光蛋白绿荧光蛋白培养尾静脉注射荧光素酶小鼠体内分布国家自然科学基金　出处：《中国组织工程研究》2017年05期 　论文类型：期刊论文

【摘要】：背景:骨髓间充质干细胞大多经静脉途径输注且归巢至骨髓的比例极低,而经股骨途径输注的研究较少。目的:观察经不同途径输注的荧光素酶基因修饰的红荧光蛋白转基因骨髓间充质干细胞在体内的分布情况。方法:生物发光法观察不同数量级的荧光素酶基因修饰的骨髓间充质干细胞(5×10~6,1×10~6,1×10~5,1×10~4)在体外孔板及游离股骨内的荧光成像情况,筛选出最佳浓度。分别经小鼠股骨和尾静脉途径输注最佳浓度的荧光素酶基因修饰的骨髓间充质干细胞,通过生物发光、病理切片、流式细胞术和荧光定量PCR法观察不同时间点两组小鼠体内荧光分布及细胞数量有无差异。结果与结论:(1)荧光素酶基因修饰的骨髓间充质干细胞的荧光强度:在体外,荧光素酶基因修饰的骨髓间充质干细胞荧光强度与细胞浓度成正相关关系;在体内,细胞输注后股骨组荧光细胞首先出现在股骨,后迅速扩散至肺,尾静脉组荧光细胞输注后迅速扩散至肺,24 h后两组荧光细胞均在脾脏、肝脏等处可见;(2)细胞分布和迁移情况:生物发光法能够实现细胞在小鼠体内分布和迁移的活体观察,细胞输注后5 min,在两组小鼠的肺脏处开始出现荧光,15 min后双肺荧光强度达到高峰,24 h后可扩散至肝、脾等组织。经股骨途径与尾静脉途径输注的骨髓间充质干细胞在小鼠体内的分布差异无显著性意义;(3)结果证实,经骨髓腔注射和尾静脉注射均不能促进骨髓间充质干细胞归巢至骨髓。
[Abstract]:Background: most of the bone marrow mesenchymal stem cells (BMSCs) are infused via vein and the proportion of homing to bone marrow is very low. Objective: to observe the distribution of erythrofluorescence gene modified bone marrow mesenchymal stem cells (BMSCs) modified by luciferase gene in vivo. Observation of bone marrow mesenchymal stem cells (BMSCs) modified by luciferase gene with different orders of magnitude by bioluminescence. 5 脳 10 ~ (6). Fluorescence imaging of 1 脳 10 ~ (6) and 1 脳 10 ~ (5) ~ (1 脳 10 ~ (4)) in external foramen plate and free femur. The best concentration was obtained. The best concentration of luciferase gene modified bone marrow mesenchymal stem cells was infused through femur and tail vein of mice respectively, and then the bone marrow mesenchymal stem cells were stained by bioluminescence and pathology. Flow cytometry and fluorescence quantitative PCR were used to observe the difference of fluorescence distribution and cell number between the two groups at different time points. Fluorescent intensity of luciferase gene modified bone marrow mesenchymal stem cells: in vitro. The fluorescence intensity of mesenchymal stem cells modified with luciferase gene was positively correlated with cell concentration. In vivo, the fluorescence cells of the femur group first appeared in the femur and then spread to the lung after the infusion of the cells, and the fluorescence cells of the tail vein group spread to the lung after 24 hours. Liver and other places can be seen; (2) Cell distribution and migration: in vivo observation of cell distribution and migration in mice by bioluminescent method. Fluorescence began to appear in the lungs of the two groups at 5 min after cell infusion. After 15 min, the fluorescence intensity of both lungs reached the peak at 24 h and then spread to the liver. There was no significant difference in the distribution of bone marrow mesenchymal stem cells between femur and caudal vein in mice. The results showed that neither medullary injection nor caudal vein injection could promote the homing of bone marrow mesenchymal stem cells to bone marrow.
【作者单位】：南方医科大学南方医院(第一临床医学院);南方医科大学南方医院血液科;广州市红十字会医院输血科;
【基金】：国家自然科学基金(30901367) 南方医院2010年度高层次课题匹配经费(20100037) 南方医科大学“优青”计划基金(C1031694)~~
【分类号】：R329.2
【正文快照】： 0引言Introduction 骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)能够分泌大量的免疫调节、抗炎、抗凋亡、促血管生成及趋化因子等生物活性分子[1-5],因此在骨髓移植相关的疾病中有广泛的用途,一方面在骨髓微环境中支持造血;另一方面,对移植免疫具有调节作用

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