携带人肝细胞生长因子的腺病毒转染人骨髓间充质干细胞及超顺磁性氧化铁标记细胞体外MR成像

发布时间：2018-01-02 12:07

本文关键词：携带人肝细胞生长因子的腺病毒转染人骨髓间充质干细胞及超顺磁性氧化铁标记细胞体外MR成像　出处：《中国医学影像技术》2013年08期 　论文类型：期刊论文

【摘要】：目的评价携带人源化绿色荧光蛋白(hrGFP)-人肝细胞生长因子(hHGF)的腺病毒载体对人永生化骨髓间充质干细胞UE7T-13生物学特征的影响,并探讨对超顺磁性氧化铁(SPIO)标记细胞进行体外MR成像的可行性。方法构建和包装携带hrGFP-hHGF基因的腺病毒载体;以hrGFP-hHGF腺病毒转染UE7T-13细胞,检测细胞中hrGFP表达阳性率、hHGF mRNA及细胞上清液中hHGF水平;检测hrGFP-hHGF腺病毒对H2O2诱导细胞凋亡的影响。以SPIO标记UE7T-13细胞,检测标记后细胞内铁含量、细胞增殖及分化能力;对不同铁浓度SPIO标记的细胞行MRI。结果成功构建hrGFP-hHGF腺病毒载体,将其转染UE7T-13细胞48h后hrGFP阳性表达率达93.17%,hHGF mRNA表达提高3075.63倍,细胞上清液中hHGF水平显著升高,随后下降,于第14天仍高于hrGFP腺病毒转染细胞。hrGFP-hHGF腺病毒可抑制H2O2介导的细胞凋亡。SPIO标记后细胞铁染色阳性率达100%,细胞铁含量明显高于未标记细胞;SPIO标记不影响细胞增殖及分化能力;T2WI信号随标记铁浓度增高而降低。结论hrGFP-hHGF腺病毒载体可抑制H2O2介导的细胞凋亡;SPIO能高效标记细胞,不影响细胞增殖及分化能力,可用于细胞体外MR成像。
[Abstract]:Objective to evaluate the expression of human green fluorescent protein (GFP) and human hepatocyte growth factor (hHGF). The effect of adenovirus vector on the biological characteristics of human immortalized bone marrow mesenchymal stem cells (UE7T-13). To explore the feasibility of in vitro Mr imaging of superparamagnetic ferric oxide labeled cells. Methods the adenovirus vector carrying hrGFP-hHGF gene was constructed and packaged. UE7T-13 cells were transfected with hrGFP-hHGF adenovirus to detect the positive rate of hrGFP expression and the level of hHGF in the supernatant. The effect of hrGFP-hHGF adenovirus on H 2O 2 induced apoptosis was detected. UE7T-13 cells were labeled with SPIO, and the iron content, proliferation and differentiation ability of UE7T-13 cells were detected. SPIO labeled cells with different iron concentrations were treated with MRI.The results showed that the adenovirus vector of hrGFP-hHGF was successfully constructed. The positive rate of hrGFP expression reached 93.17hHGF mRNA increased 3075.63 times after transfection of UE7T-13 cells for 48 hours. The level of hHGF in the supernatant increased significantly and then decreased. On the 14th day, the positive rate of iron staining of HGFP-hHGF adenovirus transfected with hrGFP was 100% after inhibiting H _ 2O _ 2-mediated apoptosis. The iron content of cells was significantly higher than that of unlabeled cells. SPIO labeling did not affect cell proliferation and differentiation. The signal intensity of T2WI decreased with the increase of labeled iron concentration. Conclusion hrGFP-hHGF adenovirus vector can inhibit the apoptosis induced by H _ 2O _ 2. SPIO can effectively label cells, without affecting cell proliferation and differentiation, and can be used for in vitro Mr imaging of cells.
【作者单位】：广州市第一人民医院放射科;中山大学附属第三医院放射科;
【基金】：国家自然科学基金(30770628、81070349) 广东省自然科学基金面上项目(8151008901000066) 广州市医药卫生科技项目(20131A011022)
【分类号】：R329
【正文快照】： 间充质干细胞(mesenchymal stromal cells,MSC)是具有自我更新和多向分化潜能的成体干细胞,不表达主要组织相容性复合体Ⅱ类分子,免疫原性低,可被受体耐受,异体移植亦无明显免疫排斥反应,为细胞移植、基因治疗以及组织器官构建的理想种子细胞[1-2],用于治疗肝纤维化具有广阔

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