Prolonged Three-Dimensional Co-Delivery of Yamanaka Factors for Cell Reprogramming.
Prolonged Three-Dimensional Co-Delivery of Yamanaka Factors for Cell Reprogramming.
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DOI：
10.1021/acsami.6b05825
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发表时间：
2016-07
影响因子：
9.5
通讯作者：
Wenwen Deng;Xia Cao;Qiang Wang;Yan Wang;Jingjing Chen;Qingtong Yu;Zhijian Zhang;Jie Zhou;Wen-qian Xu;Pan Du;Jiaxin Chen;Xiangdong Gao;Jiangnan Yu;Ximing Xu
中科院分区：
文献类型：
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作者：
Wenwen Deng;Xia Cao;Qiang Wang;Yan Wang;Jingjing Chen;Qingtong Yu;Zhijian Zhang;Jie Zhou;Wen-qian Xu;Pan Du;Jiaxin Chen;Xiangdong Gao;Jiangnan Yu;Ximing Xu
将体细胞重编程为多能状态已在二维（2D）系统中得到广泛研究，但在生物学上更忠实的三维（3D）支架中尚未描述。在这里，我们设计了一个三维多孔组织工程支架，可以实现成功和有效的诱导多能性。为了构建这种3D支架，预先通过使用磷酸钙和阳离子化的Pleuerynaeus多糖来共递送质粒OCT 4、SOX 2、KLF 4和C-MYC（pOSKM）来制备非病毒混合纳米颗粒。然后将这些混合纳米颗粒加载到3D多孔胶原支架中，以获得所谓的pOSKM活化的3D支架。这种3D支架可以将人脐带间充质干细胞（HUMSC）重编程为多能状态，产生3D细胞球，其在3D支架中显示多能性标志物的阳性表达，并且当转移到2D饲养层时紧密堆积的集落。除了与胚胎干细胞具有相似的形态学，表观遗传修饰和多能性基因表达外，3D系统生成的集落还可以在饲养层上扩增超过20代，表明成功建立了稳定的诱导多能干细胞（iPSC）系。我们的研究结果代表了首次使用多孔3D支架通过一次性转染实现成功的重编程，为iPSC生成提供了安全，简单和有效的替代策略。
Reprogramming somatic cells into a pluripotent state has been widely investigated in two-dimensional (2D) systems but not described in the more biologically faithful three-dimensional (3D) scaffolds. Here, we devise a 3D porous tissue engineering scaffold that could achieve successful and efficient induction of pluripotency. To construct this 3D scaffold, nonviral hybrid nanoparticles were fabricated beforehand by employing calcium phosphate and cationized Pleurotus eryngii polysaccharide to codeliver plasmids OCT4, SOX2, KLF4 ,and C-MYC (pOSKM). These hybrid nanoparticles were then loaded into a 3D porous collagen scaffold to obtain the so-called pOSKM-activated 3D scaffold. This 3D scaffold could reprogram human umbilical cord mesenchymal stem cells (HUMSCs) into a pluripotent state, generating 3D cell spheres which showed positive expression of pluripotency markers in the 3D scaffolds and tightly packed colonies when transferred to 2D feeder layers. Besides sharing similar morphology, epigenetic modification, and expression of pluripotency genes with the embryonic stem cells, the 3D system-generated colonies could also be expanded on feeder layers for more than 20 passages, indicating the successful establishment of stable induced pluripotent stem cell (iPSC) lines. Our findings represent a first employment of porous 3D scaffolds to achieve successful reprogramming via a one-time transfection, offering a safe, simple, and effective alternative strategy for iPSC generation.