中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (5): 676-681.doi: 10.3969/j.issn.2095-4344.2017.05.004

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

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

孙晓伟1，黄  皓2，3，周勇军2，陈晓丽1，乔鹏鑫1，邹  淳1，张秋霞2，江千里2   

  1. 1南方医科大学南方医院(第一临床医学院)，2012级临床本科在读，广东省广州市  510515；
    2南方医科大学南方医院血液科，广东省广州市  510515；
    3广州市红十字会医院输血科，广东省广州市  510220
  • 出版日期:2017-02-18 发布日期:2017-03-20
  • 通讯作者: 江千里，博士，副主任医师，南方医科大学南方医院血液科，广东省广州市 510515
  • 作者简介:孙晓伟，男，1993年生，广东省人，汉族。
  • 基金资助:

    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)

In vivo distribution of luciferase gene-labeled bone marrow mesenchymal stem cells infused through different approaches

Sun Xiao-wei1, Huang Hao2, 3, Zhou Yong-jun2, Chen Xiao-li1, Qiao Peng-xin1, Zou Chun1, Zhang Qiu-xia2, Jiang Qian-li2   

  1. 1Undergraduate of Grade 2012, First Clinical Medical College of Southern Medical University (Nanfang Hospital), Guangzhou 510515, Guangdong Province, China;
    2Department of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China;
    3Department of Blood Transfusion, Guangzhou Red Cross Hospital, Guangzhou 510220, Guangdong Province, China
  • Online:2017-02-18 Published:2017-03-20
  • Contact: Jiang Qian-li, M.D., Associate chief physician, Department of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
  • About author:Sun Xiao-wei, Undergraduate of Grade 2012, First Clinical Medical College of Southern Medical University (Nanfang Hospital), Guangzhou 510515, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 30901367; the grant from the Nanfang Hospital in 2010, No. 20100037; the Excellent Youth Fund of Southern Medical University, No. C1031694

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178

被引次数

3

摘要：

文章快速阅读：

文题释义：
荧光蛋白：运用荧光蛋白可以观测到细胞的活动，可以标记表达蛋白，可以进行深入的蛋白质组学实验等。
红色荧光蛋白：是生物学家从珊瑚虫中克隆的一种与绿色荧光蛋白同源的荧光蛋白，在紫外光的照射下可发出红色的荧光。

 

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(5×106，1×106，1×105，1×104)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。

 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID:0000-0002-2504-4697(江千里)

关键词: 干细胞, 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

Abstract:

BACKGROUND: Most bone marrow mesenchymal stem cells are infused intravenously and have very low efficiency of homing to the bone marrow. However, cell infusion via the femoral approach is little reported.
OBJECTIVE: To explore the distribution of luciferase gene modified red fluorescent protein transgenic bone marrow mesenchymal stem cells in vivo through different infusion routes.
METHODS: Luciferase gene modified bone marrow mesenchymal stem cells at different gradients (5×106, 1×106, 1×105, 1×104) were seeded or injected into the in vitro pore plate or free femurs to observe the fluorescence imaging and select the best concentration of cells. Luciferase gene modified bone marrow mesenchymal stem cells at the best cell concentration were injected into the mice via the femur and the tail vein, respectively. The distribution of fluorescence and cell number in the mice were explored by using bioluminescence, pathological examination, flow cytometry and quantitative PCR.
RESULTS AND CONCLUSION: Ex vivo fluorescence intensity of luciferase gene modified bone marrow mesenchymal stem cells was positively correlated with the cell concentration; fluorescent cells in vivo appeared in the femur first and then quickly spread to the lungs in the femur group, while fluorescent cells in the tail vein group spread to the lungs quickly after cell infusion. Fluorescent cells could be seen in the spleen, liver and other organs 24 hours later in the two groups. The distribution and migration of cells in mice could be observed successfully by bioluminescence; 5 minutes after cell infusion, the lungs of mice in the two groups began to emit fluorescence that could spread to the liver, spleen and other tissues 24 hours later, and the fluorescence intensity reached its peak after 15 minutes. The distribution of bone marrow mesenchymal stem cells in mice had no significant difference between the femur group and the tail vein group. To conclude, cell injection through the bone marrow cavity and tail vein fails to promote the homing of bone marrow mesenchymal stem cells to the bone marrow.

 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words:  Tissue Engineering, Stem Cells, Femur

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