Improved Sendai viral system for reprogramming to naive pluripotency.
Improved Sendai viral system for reprogramming to naive pluripotency.
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DOI：
10.1016/j.crmeth.2022.100317
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发表时间：
2022-11-21
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可以通过用仙台病毒（SeV）载体重编程体细胞来产生幼稚的人诱导多能干细胞（iPSC）。然而，只有真皮成纤维细胞已经成功地以这种方式重新编程，并且该过程需要在饲养细胞上培养。此外，SeV载体是高度持久的，并抑制iPSC的后续分化。在这里，我们报告了一种改良的SeV载体系统，以产生具有上级分化潜力的无转基因幼稚人iPSC。该方法不仅适用于成纤维细胞，也适用于其他类型的体细胞。SeV载体在早期传代时迅速消失，并且这种方法使得能够在无饲养层培养物中产生幼稚iPSC。通过这种方法产生的幼稚iPSC显示出比通过常规方法衍生的那些更好的分化为三系和胚胎外滋养外胚层。这种方法可以扩大iPSCs在人类早期发育和再生医学研究中的应用。一种用于产生人类iPSC的方法，其允许快速移除SeV载体。幼稚iPSC可由真皮成纤维细胞或外周血单核细胞制成。所述方法使得能够由真皮成纤维细胞产生无饲养层的幼稚iPSC。通过所述方法产生的幼稚iPSC具有显著更高的分化潜能。尽管已经报道了使用SeV载体直接由体细胞建立幼稚人类iPSC的方法，它需要使用饲养细胞上的真皮成纤维细胞作为起始群体。此外，SeV载体在生成后持续存在于iPSC中，限制了它们的分化潜能。为了解决这些问题，我们开发了一种SeV载体系统，该系统能够从各种体细胞产生幼稚iPSC，包括在无饲养层条件下。此外，该方法能够在iPSC产生后快速去除SeV载体，从而产生上级分化效力。Kunitomi等人开发改进的SeV载体系统，通过改变SeV载体的结构和组合，从各种体细胞产生幼稚人iPSC。该方法允许快速去除SeV载体，产生具有上级分化潜力的无转基因幼稚iPSC。
Naive human induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with Sendai virus (SeV) vectors. However, only dermal fibroblasts have been successfully reprogrammed this way, and the process requires culture on feeder cells. Moreover, SeV vectors are highly persistent and inhibit subsequent differentiation of iPSCs. Here, we report a modified SeV vector system to generate transgene-free naive human iPSCs with superior differentiation potential. The modified method can be applied not only to fibroblasts but also to other somatic cell types. SeV vectors disappear quickly at early passages, and this approach enables the generation of naive iPSCs in a feeder-free culture. The naive iPSCs generated by this method show better differentiation to trilineage and extra-embryonic trophectoderm than those derived by conventional methods. This method can expand the application of iPSCs to research on early human development and regenerative medicine. A method for human iPSC generation that allows rapid removal of SeV vector Naive iPSCs can be made from dermal fibroblasts or peripheral blood mononuclear cells The method enables feeder-free naive iPSCs generation from dermal fibroblasts The naive iPSCs generated by the method have significantly higher differentiation potency Although a method to establish naive human iPSCs directly from somatic cells using SeV vectors has been reported, it requires the use of dermal fibroblasts on feeder cells as a starting population. Furthermore, the SeV vector persists in the iPSCs after generation, limiting their differentiation potency. To solve these problems, we developed a SeV vector system that enables the generation of naive iPSCs from a variety of somatic cells, including under feeder-free conditions. Moreover, the method enables rapid removal of SeV vectors after iPSC generation, resulting in a superior differentiation potency. Kunitomi et al. develop an improved SeV vector system to generate naive human iPSCs from various somatic cells by changing the structure and combination of SeV vectors. This method allows rapid removal of the SeV vectors, resulting in transgene-free naive iPSCs with superior differentiation potential.