Direct Reprogramming of Human Suspension Cells into Mesodermal Cell Lineages via Combined Magnetic Targeting and Photothermal Stimulation by Magnetic Graphene Oxide Complexes
Direct Reprogramming of Human Suspension Cells into Mesodermal Cell Lineages via Combined Magnetic Targeting and Photothermal Stimulation by Magnetic Graphene Oxide Complexes
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DOI：
10.1002/smll.201700703
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发表时间：
2017-08-25
期刊：
影响因子：
13.3
通讯作者：
Chen, San-Yuan
中科院分区：
文献类型：
--
作者：
Chiang, Min-Yu;Lin, Yi-Zhen;Chen, San-Yuan
悬浮细胞可以为细胞重编程提供细胞来源，但它们很难通过非病毒载体转染。以氧化铁纳米颗粒(Fe3O4)修饰的氧化石墨烯(GO)与聚乙烯亚胺(PEI)复合，构建了一种高效、安全的非病毒载体(GO-Fe3O4-PEI复合体)，可同时将编码Oct3/4、针对P53、Sox2、Klf4、L-Myc和Lin28的多能相关因子的3种异构体载体(pCXLE-hOCT3/4-shp53、pCXLE-HSK和pCXLE-HUL)同时导入人外周血单核细胞(PBMC)。采用磁力搅拌和近红外(NIR)激光照射相结合的方法，分别促进复合体与悬浮细胞之间的接触，提高细胞膜的通透性，对悬浮PBMCs进行多种物理刺激。聚合酶链式反应分析表明，磁靶向和光热刺激的联合作用明显提高了悬浮细胞的转染率。转基因细胞表达的多能性标志物包括Nanog、Oct4和Sox2，具有向中胚层和外胚层细胞分化的潜能。结果表明，GO-Fe3O4-PEI复合体为将PBMC重编程为部分诱导的多能干细胞提供了一种安全、方便和有效的工具，无需完全重编程即可快速转分化为中胚层细胞。
Suspension cells can provide a source of cells for cellular reprogramming, but they are difficult to transfect by nonviral vectors. An efficient and safe nonviral vector (GO-Fe3O4-PEI complexes) based on iron oxide nanoparticle (Fe3O4)-decorated graphene oxide (GO) complexed with polyethylenimine (PEI) for the first time is developed for delivering three individual episomal plasmids (pCXLE-hOCT3/4-shp53, pCXLE-hSK, and pCXLE-hUL) encoding pluripotent-related factors of Oct3/4, shRNA against p53, Sox2, Klf4, L-Myc, and Lin28 into human peripheral blood mononuclear cells (PBMCs) simultaneously. The combined treatment of magnetic stirring and near-infrared (NIR)-laser irradiation, which can promote contact between the complexes and floating cells and increase the cell membrane permeability, respectively, is used to conduct multiple physical stimulations for suspension PBMCs transfection. The PCR analysis shows that the combinatorial effect of magnetic targeting and photothermal stimulation obviously promoted the transfection efficiency of suspension cells. The transfected cells show positive expression of the pluripotency markers, including Nanog, Oct4, and Sox2, and have potential to differentiate into mesoderm and ectoderm cells. The results demonstrate that the GO-Fe3O4-PEI complex provides a safe, convenient, and efficient tool for reprogramming PBMCs into partially induced pluripotent stem cells, which are able to rapidly transdifferentiate into mesodermal lineages without full reprogramming.