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利用纳米材料介导的药物靶向治疗和动物细胞转基因等相关研究,日益受到人们的关注.但植物因存在细胞壁的障碍,无论原位还是离体细胞培养条件下,利用纳米技术进行基因转移均存在很大难度.因此设想,如通过纳米颗粒材料物理尺寸的改变和表面化学修饰,能改变纳米颗粒与植物细胞壁界面上的生物物理或生物化学特征,从而有利于纳米颗粒材料穿越植物细胞壁进入植物细胞,将对推动纳米技术在植物转基因领域中的应用产生重要意义.根据以上设想,研究了不同的共孵育时间和温度等条件下,杂交鹅掌楸的胚性悬浮细胞与经不同表面化学修饰的CdSe/ZnS纳米颗粒之间相互作用过程的细胞生物学特征,以及CdSe/ZnS量子点的细胞毒性.结果表明,在共孵育后3h以内,激光共聚焦显微镜和电子扫描显微镜下,均可观察到经表面后修饰带正电荷的CdSe/ZnS纳米颗粒.同时,胞吞进入细胞内部的表面携带正电荷的CdSe/ZnS纳米颗粒的量明显与共培养时间、温度有明显的依赖关系,表明它们可以通过细胞的液相胞吞作用进入杂交鹅掌楸细胞内,且不影响细胞的活性;而表面带负电荷的CdSe/ZnS纳米颗粒则主要聚集在细胞外壁附近.在培养溶液中添加20%(质量比)聚乙二醇,可进一步提高鹅掌楸细胞胞吞CdSe/ZnS纳米颗粒的量和减轻CdSe/ZnS纳米颗粒的细胞毒性.本研究表明,以表面携带正电荷的CdSe/ZnS量子点纳米材料作为基因载体,在植物悬浮细胞的转基因研究和应用中具有广泛的前景.
The use of nanomaterial-mediated drug-targeted therapies and animal cell transgenesis and other related research, more and more people’s attention.But plants due to the presence of cell wall barriers, either in situ or in vitro cell culture conditions, the use of nanotechnology for gene transfer are Therefore, it is envisaged that the biophysical or biochemical characteristics at the interface between the nanoparticle and the cell wall of a plant may be changed by changing the physical size of the nanoparticle material and modifying the surface chemical, so as to facilitate the crossing of the nanoparticle material through the plant cell wall into the plant Cells will play an important role in promoting the application of nanotechnology in the field of plant genetics.Based on the above assumptions, under different conditions of co-incubation time and temperature, embryogenic suspension cells of hybrid Liriodendron hybridisation with different surface chemical modification Of CdSe / ZnS nanoparticles and the cytotoxicity of CdSe / ZnS quantum dots.The results showed that within 3h after co-incubation, the results of confocal laser scanning microscopy and scanning electron microscopy After surface modification, positively charged CdSe / ZnS nanoparticles were modified The amount of positively charged CdSe / ZnS nanoparticles on the inner surface of cells was obviously dependent on the time and temperature of co-culture, indicating that they could enter the cells of hybrid Liriodendron by liquid-phase endocytosis without affecting the cell While the negatively charged CdSe / ZnS nanoparticles mainly gathered near the outer wall of the cells.Adding 20% (mass ratio) polyethylene glycol to the culture solution could further enhance the cell growth of CdSe / ZnS The amount of nanoparticles and mitigating the cytotoxicity of CdSe / ZnS nanoparticles.The present study shows that CdSe / ZnS quantum dot nanomaterials with positive charges on the surface as gene carriers have a wide range of prospects in the study and application of transgenic plant cells in suspension .