Aim: In this study, we aimed to develop a polycationic non-viral carrier for the delivery of the reprogramming factors to the L929 fibroblast cell.Methods: We have prepared (3-hydroxybutyrate-co-3-hydroxyhexanoate) PHBHHx-based nanoparticles with the solvent diffusion method. Cytotoxicity of PXNs was determined via MTT assay. Transfection efficiency was evaluated via screening GFP expression by fluorescence microscopy. The expression of reprogramming factors (Oct4, Klf4, and Sox2) was determined by RT-qPCR.Results: PXNs with 32.9 ± 0.41 mV zeta potential and 177.6 ± 0.80 nm size were used for transfection of L929 Fbroblast cells. The percentage of cell viability of PXN were between 91.8%(±2.9) and 42.1%(±1.3). The transfection efficiency was found as 71.6%(±3,5). According to RT-qPCR data, the rate of transfection factors was significantly increased after the 11th cycle compared to non-transfected cells. Based on these results, it can be concluded that newly developed PXN is thought to be an effective tool for reprogramming cells.

中文翻译：

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基于3-羟基己酸酯的聚阳离子纳米粒子系统，用于传递重编程因子。

目的：在这项研究中，我们旨在开发一种聚阳离子非病毒载体，用于将重编程因子递送至L929成纤维细胞。方法：我们制备了（3-羟基丁酸酯-co-3-羟基己酸酯）基于PHBHHx的纳米颗粒溶剂扩散法。通过MTT测定法测定PXN的细胞毒性。通过荧光显微镜检查GFP表达来评估转染效率。通过RT-qPCR确定重编程因子（Oct4，Klf4和Sox2）的表达。结果：将具有32.9±0.41mV zeta电位和177.6±0.80nm大小的PXN用于转染L929成纤维细胞。PXN的细胞存活率百分比在91.8％（±2.9）和42.1％（±1.3）之间。转染效率为71.6％（±3,5）。根据RT-qPCR数据，与未转染的细胞相比，第11个周期后转染因子的比率显着增加。根据这些结果，可以得出结论，新开发的PXN被认为是对细胞进行重新编程的有效工具。