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ScreenFect A:高效且低毒的脂質體,轉染間充質干細胞

更新時間:2023-04-01   點擊次數:374次

ScreenFect A: an efficient and low toxic liposome for gene delivery to mesenchymal stem cells

ScreenFect A:一種高效且低毒的脂質體,用于將基因傳遞到間充質干細胞

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摘要

間充質干細胞(MSCs)在包括組織工程和癌癥治療在內的各種治療應用中具有巨大的前景。間充質干細胞的基因修飾可用于通過促進特定功能或將間充質干細胞轉化為更有效的基因治療工具來增強間充質干細胞的治療效果。然而,轉基因MSC的成功生成往往受到現有轉染試劑轉染效率差或毒性高的限制。在我們之前的研究中,我們使用硫醇-炔點擊化學來開發新的脂質體載體,包括ScreenFect(A)  (® SF)(Li等人,2012)。本研究考察了SF對MSCs的轉染性能。以Lipofectamine™ 2000(L2K)為對照,對轉染效率、細胞活力和細胞DNA攝取進行了比較評估,結果表明SF在MSC轉染方面優于L2K。血清的存在對SF或L2K的轉染效率沒有顯著影響,但大大降低了L2K轉染MSC的活力。與L2K相比,SF介導的轉染效率更高,也與更好的細胞增殖相關。這些結果得到了DNA細胞內命運監測的支持,這證實了DNA從溶酶體的穩定運輸和有效的核定位。SF的TGF-β1基因遞送促進了成骨誘導條件下MSC成骨分化。作為SF脂質化對干細胞的研究,本研究強調了SF在向間充質干細胞和其他干細胞的基因傳遞中促進基于轉基因干細胞的組織工程和其他治療效果的有希望的作用。


Abstract

Mesenchymal stem cells (MSCs) hold great promise in variety of therapeutic applications including tissue engineering and cancer therapy. Genetic modification of MSCs can be used to enhance the therapeutic effect of MSCs by facilitating a specific function or by transforming MSCs into more effective gene therapy tools. However, the successful generation of genetically modified MSCs is often limited by the poor transfection efficiency or high toxicity of available transfection reagents. In our previous study, we used thiol-yne click chemistry to develop new liposomal vectors, including ScreenFect(®) A (SF) (Li et al., 2012). In this study, we investigated the transfection performance of SF on MSCs. A comparative evaluation of transfection efficiency, cell viability and cellular DNA uptake was performed using the Lipofectamine™ 2000 (L2K) as a control, and the results show that SF is superior to L2K for MSC transfection. The presence of serum did not significantly influence the transfection efficiency of either SF or L2K but greatly reduced the viability of MSC transfected by L2K. The higher efficiency of SF-mediated transfection compared to L2K was also correlated with better proliferation of cells. These results were supported by monitoring the intracellular fate of DNA, which confirmed stable transportation of DNA from lysosomes and efficient nuclear localization. TGF-β1 gene delivery by SF promoted MSC osteogenic differentiation in an osteogenic induction condition. As the first study of SF lipofection on stem cells, this study highlights a promising role of SF in gene delivery to MSCs as well as other stem cells to facilitate tissue engineering and other therapeutic effects based on genetically modified stem cells.


通過科研成果轉化,ScreenFect已經成為一種商業轉染試劑。德國ScreentFect聯合靶點科技,將這種通過點擊化學篩選出來的高效轉染試劑帶給中國廣大的科研工作者。詳情聯系靶點科技。


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