Department of Soft Condensed Matter, Institute of Physical Chemistry, Polish Academy of Sciences, 01-224 Warsaw, Poland.
J Colloid Interface Sci. 2022 Dec;627:270-282. doi: 10.1016/j.jcis.2022.07.040. Epub 2022 Jul 9.
Most experimental procedures applied in modern biology involve cargo delivering into cells. One of the ways to cargo introduction is osmotic-mediated intracellular vesicle swelling. However, its widespread use was hindered due to cargo size (<10 nm) and cell-type-related restrictions. We addressed the issue of the composition of colloidal loading solution to enhance the efficiency of cellular delivery.
We examined the effectiveness of colloidal loading solutions of varied compositions, including various types and sizes of polymers building osmotic pressure. We used confocal imaging coupled with fluorescence correlation spectroscopy to evaluate the introduction of polymers, proteins, nanoparticles, and DNA plasmids (cargos of sizes 1-175 nm) to cells representing eight cell lines: cancer, normal, epithelial, and mesenchymal ones.
We found that cellular delivery effectiveness strongly correlates with the size and concentration of osmotic pressure building polymers and not with the high value of the osmotic pressure itself. We show that polymer solutions at the entangled regime of concentrations enhance the delivery of large biomacromolecules even of size 200 nm (DNA plasmids) into cells, including MDA-MB-231 cells - so far resistant to the osmotic procedure. We show that the colloid loading medium based on entangled polymer chains is a versatile cargo delivery tool for molecular biology.
大多数应用于现代生物学的实验程序都涉及将货物输送到细胞中。将货物引入细胞的一种方法是通过渗透压介导的细胞内囊泡肿胀。然而，由于货物尺寸(<10nm)和细胞类型相关的限制，其广泛应用受到阻碍。我们解决了胶体加载溶液的组成问题，以提高细胞输送的效率。
我们研究了不同组成的胶体加载溶液的有效性，包括构建渗透压的各种类型和大小的聚合物。我们使用共聚焦成像和荧光相关光谱技术来评估聚合物、蛋白质、纳米颗粒和 DNA 质粒（尺寸为 1-175nm 的货物）在代表八种细胞系（癌症、正常、上皮和间充质）的细胞中的引入情况。
我们发现细胞输送效率与构建渗透压的聚合物的尺寸和浓度强烈相关，而与渗透压本身的高值无关。我们表明，聚合物溶液在浓度的缠结状态下增强了大生物大分子的输送，即使是尺寸为 200nm 的 DNA 质粒（DNA 质粒）也能进入细胞，包括迄今为止对渗透压程序有抗性的 MDA-MB-231 细胞。我们表明，基于缠结聚合物链的胶体加载介质是分子生物学中一种通用的货物输送工具。
