Non-Viral Co-Delivery of the Four Yamanaka Factors for Generation of Human Induced Pluripotent Stem Cells via Calcium Phosphate Nanocomposite Particles
Non-Viral Co-Delivery of the Four Yamanaka Factors for Generation of Human Induced Pluripotent Stem Cells via Calcium Phosphate Nanocomposite Particles
复制标题
通过磷酸钙纳米复合颗粒非病毒共同递送四种山中因子以产生人类诱导多能干细胞
DOI：
10.1002/adfm.201203646
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发表时间：
2013-11-20
影响因子：
19
通讯作者：
Yu, Jiangnan
中科院分区：
文献类型：
--
作者：
Cao, Xia;Deng, Wenwen;Yu, Jiangnan
诱导多能干细胞（iPSC）的产生可以通过特定转录因子集的异位表达来实现。然而，大多数iPSC诱导的实例都是使用病毒载体实现的，这带来了不可预测的遗传功能障碍的风险。在这里，首次报道了一种基于磷酸钙纳米颗粒的非病毒载体，用于通过共递送四种质粒（Oct 4，Sox 2，Klf 4和c-Myc）从人脐带间充质干细胞（HUMSC）产生无病毒的iPSC。结果，从20万个细胞中获得了总共98个集落，重编程效率为0.049%。iPSC显示多能性标志物的阳性表达，所述多能性标志物包括0 CT 4、SSEA-3、SSEA-4、NANOG和TRA-1-81。此外，iPSC能够在体外分化成所有三个胚层。将iPSC皮下注射到免疫功能低下的小鼠中导致形成含有来自所有三个胚层的各种组织的畸胎瘤。这些发现表明，通过质粒包裹的磷酸钙纳米颗粒共递送四种Yamanaka因子可以为无病毒iPSCs的产生提供简单，安全和有效的方法，这对于它们未来在再生医学领域的临床应用至关重要。
Generating of induced pluripotent stem cells (iPSCs) can be achieved by ectopic expression of defined transcription factor sets. However, most instances of iPSC induction have been achieved using viral vectors, which carry the risk of unpredictable genetic dysfunction. Here, for the first time, a non-viral vector based on calcium phosphate nanoparticles for the generation of virus-free iPSCs from human umbilical cord mesenchymal stem cells (HUMSCs) via co-delivery of the four plasmids (Oct4, Sox2, Klf4, and c-Myc) is reported. As a result, a total of 98 colonies from 200 000 cells have been obtained, with a reprogramming efficiency of 0.049%. The iPSCs shows positive expression of pluripotency markers, including OCT4, SSEA-3, SSEA-4, NANOG, and TRA-1-81. Moreover, the iPSCs are able to differentiate into all three germ layers in vitro. Subcutaneous injection of the iPSCs into immunocompromised mice results in the formation of teratomas containing a variety of tissues from all three germ layers. These findings indicate that co-delivery of the four Yamanaka factors via plasmid-encapsulated calcium phosphate nanoparticles can provide a simple, safe, and efficient method for the generation of virus-free iPSCs, which is crucial for their future clinical applications in the field of regenerative medicine.