Department of Biomedical Science, College of Life Science, CHA University 606-16, Yeoksam 1-Dong, Kangnam-gu, Seoul 135-081, Republic of Korea.
Biomaterials. 2011 Jan;32(1):268-78. doi: 10.1016/j.biomaterials.2010.08.086. Epub 2010 Sep 27.
In stem cell therapy, transfection of specific genes into stem cells is an important technique to induce cell differentiation. To perform gene transfection in human mesenchymal stem cells (hMSCs), we designed and fabricated a non-viral vector system for specific stem cell differentiation. Several kinds of gene carriers were evaluated with regard to their transfection efficiency and their ability to enhance hMSCs differentiation. Of these delivery vehicles, biodegradable poly (DL-lactic-co-glycolic acid) (PLGA) nanoparticles yielded the best results, as they complexed with high levels of plasmid DNA (pDNA), allowed robust gene expression in hMSCs, and induced chondrogenesis. Polyplexing with polyethylenimine (PEI) enhanced the cellular uptake of SOX9 DNA complexed with PLGA nanoparticles both in vitro and in vivo. The expression of enhanced green fluorescent protein (EGFP) and SOX9 increased up to 75% in hMSCs transfected with PEI/SOX9 complexed PLGA nanoparticles 2 days after transfection. SOX9 gene expression was evaluated by RT-PCR, real time-qPCR, glycosaminoglycan (GAG)/DNA levels, immunoblotting, histology, and immunofluorescence.
在干细胞治疗中，将特定基因转染到干细胞中是诱导细胞分化的重要技术。为了在人骨髓间充质干细胞（hMSCs）中进行基因转染，我们设计并制造了一种用于特定干细胞分化的非病毒载体系统。我们评估了几种基因载体的转染效率及其增强 hMSCs 分化的能力。在这些递送载体中，可生物降解的聚（DL-乳酸-共-乙醇酸）（PLGA）纳米粒的效果最佳，因为它们可以与高水平的质粒 DNA（pDNA）复合，在 hMSCs 中产生强大的基因表达，并诱导软骨生成。与聚乙烯亚胺（PEI）复合可增强 SOX9 与 PLGA 纳米粒复合物在体外和体内的 hMSCs 摄取。转染 PEI/SOX9 复合 PLGA 纳米粒 2 天后，hMSCs 中增强型绿色荧光蛋白（EGFP）和 SOX9 的表达增加了 75%。通过 RT-PCR、实时 qPCR、糖胺聚糖（GAG）/DNA 水平、免疫印迹、组织学和免疫荧光评估 SOX9 基因表达。
