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ScreenFect A:高效且低毒的脂質(zhì)體,轉(zhuǎn)染間充質(zhì)干細(xì)胞

更新時(shí)間:2023-04-01   點(diǎn)擊次數(shù):1034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(zhì)體,用于將基因傳遞到間充質(zhì)干細(xì)胞

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摘要

間充質(zhì)干細(xì)胞(MSCs)在包括組織工程和癌癥治療在內(nèi)的各種治療應(yīng)用中具有巨大的前景。間充質(zhì)干細(xì)胞的基因修飾可用于通過(guò)促進(jìn)特定功能或?qū)㈤g充質(zhì)干細(xì)胞轉(zhuǎn)化為更有效的基因治療工具來(lái)增強(qiáng)間充質(zhì)干細(xì)胞的治療效果。然而,轉(zhuǎn)基因MSC的成功生成往往受到現(xiàn)有轉(zhuǎn)染試劑轉(zhuǎn)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(diǎn)擊化學(xué)來(lái)開(kāi)發(fā)新的脂質(zhì)體載體,包括ScreenFect(A)  (® SF)(Li等人,2012)。本研究考察了SF對(duì)MSCs的轉(zhuǎn)染性能。以Lipofectamine™ 2000(L2K)為對(duì)照,對(duì)轉(zhuǎn)染效率、細(xì)胞活力和細(xì)胞DNA攝取進(jìn)行了比較評(píng)估,結(jié)果表明SF在MSC轉(zhuǎn)染方面優(yōu)于L2K。血清的存在對(duì)SF或L2K的轉(zhuǎn)染效率沒(méi)有顯著影響,但大大降低了L2K轉(zhuǎn)染MSC的活力。與L2K相比,SF介導(dǎo)的轉(zhuǎn)染效率更高,也與更好的細(xì)胞增殖相關(guān)。這些結(jié)果得到了DNA細(xì)胞內(nèi)命運(yùn)監(jiān)測(cè)的支持,這證實(shí)了DNA從溶酶體的穩(wěn)定運(yùn)輸和有效的核定位。SF的TGF-β1基因遞送促進(jìn)了成骨誘導(dǎo)條件下MSC成骨分化。作為SF脂質(zhì)化對(duì)干細(xì)胞的研究,本研究強(qiáng)調(diào)了SF在向間充質(zhì)干細(xì)胞和其他干細(xì)胞的基因傳遞中促進(jìn)基于轉(zhuǎn)基因干細(xì)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(guò)科研成果轉(zhuǎn)化,ScreenFect已經(jīng)成為一種商業(yè)轉(zhuǎn)染試劑。德國(guó)ScreentFect聯(lián)合靶點(diǎn)科技,將這種通過(guò)點(diǎn)擊化學(xué)篩選出來(lái)的高效轉(zhuǎn)染試劑帶給中國(guó)廣大的科研工作者。詳情聯(lián)系靶點(diǎn)科技。


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