兔骨髓间充质干细胞培养及体内示踪研究

发布时间：2018-05-21 08:47

本文选题：骨髓间充质干细胞 + 多聚赖氨酸修饰的四氧化三铁磁性纳米颗粒　；参考：《昆明医科大学》2015年硕士论文

【摘要】：目的采用贴壁法体外培养扩增兔骨髓间充质干细胞(Bone Mesenchymal Stem Cells, BMSCs)。用PLL@Fe3O4对兔骨髓间充质干细胞(BMSCs)进行体外标记,将标记过的细胞经兔耳缘静脉注入,行MRI检查了解其在肝脏中的分布情况。为干细胞移植治疗、MRI活体示踪提供基础性实验数据。方法1.采用贴壁法培养扩增兔BMSCs,通过形态学观察及流式细胞仪鉴定证实得到的细胞为BMSCs。2.用PLL@Fe3O4标记BMSCs。3. PLL@Fe3O4标记过的干细胞进行普鲁士兰染色,并在显微镜下观察其在BMSCs中的标记情况。4.台盼兰排斥实验及MTT法测定PLL@Fe3O4对BMSCs增殖及其活性的影响。5.将PLL@Fe3O4标记的兔BMSCs通过耳缘静脉输入兔体内行MRI检测,观察其在肝脏中的分布情况。结果1.倒置相差显微镜可观察到大量梭形和漩涡状贴壁细胞,流式细胞仪鉴定可见所培养的间充质干细胞高表达其表面标记物CD44,低表达CD14和CD45等造血细胞表面标志,可初步证实所培养的细胞为兔BMSCs。2.20 μg/ml PLL@Fe3O4标记的兔BMSCs对细胞无明显损伤及毒性,且标记率高,可安全用于细胞的标记。3.MRI检测在T2*W1序列上能明显观察到]PLL@Fe3O4标记的兔BMSCs可沉积在肝脏及其周围血管内,证明骨髓间充质于细胞可以通过外周血管进入肝脏。
[Abstract]:Objective to amplify bone Mesenchymal Stem Cells, BMSCs from rabbit bone marrow mesenchymal stem cells (BMSCs) by adherent method in vitro. Rabbit bone marrow mesenchymal stem cells (BMSCs) were labeled with PLL@Fe3O4 in vitro. The labeled cells were injected into rabbit auricular vein and the distribution of BMSCs in liver was investigated by MRI. To provide the basic experimental data for the transplantation of stem cell for MRI in vivo tracer. Method 1. Rabbit BMSCs were amplified by adherent method. The cells were identified as BMSCs.2by morphological observation and flow cytometry. PLL@Fe3O4 was used to mark BMSCs.3. PLL@Fe3O4 labeled stem cells were stained with Prussian blue and their labeling in BMSCs was observed under microscope. Trypan blue exclusion test and MTT assay were used to determine the effect of PLL@Fe3O4 on the proliferation and activity of BMSCs. Rabbit BMSCs labeled with PLL@Fe3O4 was injected into rabbit via auricular vein to detect MRI and its distribution in liver was observed. Result 1. A large number of fusiform and whirlpool adherent cells were observed under inverted phase contrast microscope. The cultured mesenchymal stem cells expressed high expression of CD44, and low expression of CD14 and CD45 markers on the surface of hematopoietic cells. It can be preliminarily proved that the cultured cells labeled with BMSCs.2.20 渭 g/ml PLL@Fe3O4 have no obvious damage and toxicity to the cells, and the labeling rate is high. PLL@Fe3O4 labeled rabbit BMSCs could be deposited in the liver and its surrounding vessels, which proved that the bone marrow mesenchymal cells could enter the liver via peripheral blood vessels.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R657.3

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