本文選題：骨髓間充質(zhì)干細胞 + 多聚賴氨酸修飾的四氧化三鐵磁性納米顆粒�。� 參考：《昆明醫(yī)科大學》2015年碩士論文

【摘要】：目的采用貼壁法體外培養(yǎng)擴增兔骨髓間充質(zhì)干細胞(Bone Mesenchymal Stem Cells, BMSCs)。用PLL@Fe3O4對兔骨髓間充質(zhì)干細胞(BMSCs)進行體外標記,將標記過的細胞經(jīng)兔耳緣靜脈注入,行MRI檢查了解其在肝臟中的分布情況。為干細胞移植治療、MRI活體示蹤提供基礎(chǔ)性實驗數(shù)據(jù)。方法1.采用貼壁法培養(yǎng)擴增兔BMSCs,通過形態(tài)學觀察及流式細胞儀鑒定證實得到的細胞為BMSCs。2.用PLL@Fe3O4標記BMSCs。3. PLL@Fe3O4標記過的干細胞進行普魯士蘭染色,并在顯微鏡下觀察其在BMSCs中的標記情況。4.臺盼蘭排斥實驗及MTT法測定PLL@Fe3O4對BMSCs增殖及其活性的影響。5.將PLL@Fe3O4標記的兔BMSCs通過耳緣靜脈輸入兔體內(nèi)行MRI檢測,觀察其在肝臟中的分布情況。結(jié)果1.倒置相差顯微鏡可觀察到大量梭形和漩渦狀貼壁細胞,流式細胞儀鑒定可見所培養(yǎng)的間充質(zhì)干細胞高表達其表面標記物CD44,低表達CD14和CD45等造血細胞表面標志,可初步證實所培養(yǎng)的細胞為兔BMSCs。2.20 μg/ml PLL@Fe3O4標記的兔BMSCs對細胞無明顯損傷及毒性,且標記率高,可安全用于細胞的標記。3.MRI檢測在T2*W1序列上能明顯觀察到]PLL@Fe3O4標記的兔BMSCs可沉積在肝臟及其周圍血管內(nèi),證明骨髓間充質(zhì)于細胞可以通過外周血管進入肝臟。
[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.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R657.3

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