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  本文關(guān)鍵詞：攜帶人肝細胞生長因子的腺病毒轉(zhuǎn)染人骨髓間充質(zhì)干細胞及超順磁性氧化鐵標(biāo)記細胞體外MR成像　出處：《中國醫(yī)學(xué)影像技術(shù)》2013年08期 　論文類型：期刊論文

  更多相關(guān)文章： 間充質(zhì)干細胞 基因轉(zhuǎn)染 肝細胞生長因子 超順磁性氧化鐵 磁共振成像

【摘要】：目的評價攜帶人源化綠色熒光蛋白(hrGFP)-人肝細胞生長因子(hHGF)的腺病毒載體對人永生化骨髓間充質(zhì)干細胞UE7T-13生物學(xué)特征的影響,并探討對超順磁性氧化鐵(SPIO)標(biāo)記細胞進行體外MR成像的可行性。方法構(gòu)建和包裝攜帶hrGFP-hHGF基因的腺病毒載體;以hrGFP-hHGF腺病毒轉(zhuǎn)染UE7T-13細胞,檢測細胞中hrGFP表達陽性率、hHGF mRNA及細胞上清液中hHGF水平;檢測hrGFP-hHGF腺病毒對H2O2誘導(dǎo)細胞凋亡的影響。以SPIO標(biāo)記UE7T-13細胞,檢測標(biāo)記后細胞內(nèi)鐵含量、細胞增殖及分化能力;對不同鐵濃度SPIO標(biāo)記的細胞行MRI。結(jié)果成功構(gòu)建hrGFP-hHGF腺病毒載體,將其轉(zhuǎn)染UE7T-13細胞48h后hrGFP陽性表達率達93.17%,hHGF mRNA表達提高3075.63倍,細胞上清液中hHGF水平顯著升高,隨后下降,于第14天仍高于hrGFP腺病毒轉(zhuǎn)染細胞。hrGFP-hHGF腺病毒可抑制H2O2介導(dǎo)的細胞凋亡。SPIO標(biāo)記后細胞鐵染色陽性率達100%,細胞鐵含量明顯高于未標(biāo)記細胞;SPIO標(biāo)記不影響細胞增殖及分化能力;T2WI信號隨標(biāo)記鐵濃度增高而降低。結(jié)論hrGFP-hHGF腺病毒載體可抑制H2O2介導(dǎo)的細胞凋亡;SPIO能高效標(biāo)記細胞,不影響細胞增殖及分化能力,可用于細胞體外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.
【作者單位】： 廣州市第一人民醫(yī)院放射科;中山大學(xué)附屬第三醫(yī)院放射科;
【基金】：國家自然科學(xué)基金(30770628、81070349) 廣東省自然科學(xué)基金面上項目(8151008901000066) 廣州市醫(yī)藥衛(wèi)生科技項目(20131A011022)
【分類號】：R329
【正文快照】： 間充質(zhì)干細胞(mesenchymal stromal cells,MSC)是具有自我更新和多向分化潛能的成體干細胞,不表達主要組織相容性復(fù)合體Ⅱ類分子,免疫原性低,可被受體耐受,異體移植亦無明顯免疫排斥反應(yīng),為細胞移植、基因治療以及組織器官構(gòu)建的理想種子細胞[1-2],用于治療肝纖維化具有廣闊

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本文編號：1369142