Polyethyleneimine-mediated gene delivery into human adipose derived stem cells
Polyethyleneimine-mediated gene delivery into human adipose derived stem cells
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DOI：
10.1016/j.biomaterials.2008.02.006
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发表时间：
2008-05-01
期刊：
影响因子：
14
通讯作者：
Lee, Hai Bang
中科院分区：
文献类型：
--
作者：
Ahn, Hyun Hee;Lee, Jung Hwa;Lee, Hai Bang
在这项研究中，我们研究了使用聚乙烯亚胺（PEI）作为人类脂肪组织来源的干细胞（hADSC）的基因递送载体。这些多能细胞可以形成骨、软骨、脂肪和其他结缔组织。在原代培养中，hADSC在原代培养中的外观是成纤维细胞，并且它们在至少五代中显示出高增殖率。免疫表型分析表明，这些细胞是阳性的间充质干细胞标志物CD 29和CD 44，但阴性的造血细胞表面标志物CD 34，CD 45，和c-kit。比较PEI和Lipofectamine作为hADSC的基因载体。DNA以4的负-正（N/P）电荷比完全结合PEI。PEI-DNA复合物呈球形，表面光滑。随着PEI比例的增加，PEI-DNA复合物的尺寸从990 nm减小到130 nm，表面正电荷减少，细胞毒性增加。流式细胞术显示，使用PEI在N/P电荷比为4和8时的转染效率高于Lipofectamine。在N/P电荷比为8时获得最高的转染效率（19%）。转染后，增强型绿色荧光蛋白（EGFP）在4 μ m ~ 30 μ m处开始定位于细胞核内，在9 μ m ~ 30 μ m处开始定位于细胞质内。总之，PEI作为hADSC的有效基因载体。(c)2008爱思唯尔有限公司保留所有权利。
In this study, we examined the use of polyethyleneimine (PEI) as a carrier for gene delivery in human adipose tissue-derived stem cells (hADSCs). These multipotent cells can form bone, cartilage, adipose, and other connective tissues. In primary culture, hADSCs are fibroblastic in appearance in primary culture, and they show a high rate of proliferation for at least five passages. Immunophenotyping showed that these cells are positive for the mesenchymal stem cell markers CD29 and CD44 but negative for the hematopoietic cell surface markers CD34, CD45, and c-kit. PEI and Lipofectamine were compared as gene carriers for hADSCs. DNA completely bound PEI at a negative-to-positive (N/P) charge ratio of 4. The PEI-DNA complexes were spherical with smooth surfaces. As the proportion of PEI was increased, the size of the PEI-DNA complexes decreased from 990 to 130 nm, the positive surface charge decreased, and the cytotoxicity increased. Flow cytometry revealed that the transfection efficiency using PEI at N/P charge ratios of 4 and 8 was higher than that of Lipofectamine. The highest transfection efficiency (19%) was obtained at an N/P charge ratio of 8. After transfection, the enhanced green fluorescent protein (EGFP) started to localize in the nuclei of hADSCs at 4 It 30 m and localize over cytoplasm from 9 It 30 m. In conclusion, PEI acts as an effective gene carrier for hADSCs. (c) 2008 Elsevier Ltd. All rights reserved.