Antibody-directed lentiviral gene transduction for live-cell monitoring and selection of human iPS and hES cells.
Antibody-directed lentiviral gene transduction for live-cell monitoring and selection of human iPS and hES cells.
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DOI：
10.1371/journal.pone.0034778
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Roth MJ
中科院分区：
文献类型：
--
作者：
Wu DT;Seita Y;Zhang X;Lu CW;Roth MJ
在混合细胞群中识别干细胞是干细胞生物学的一个主要障碍，特别是在重编程过程中识别诱导多能干细胞（iPS）。基于干细胞表面标志物的选择性表达，已经开发了通过抗体缀合的慢病毒颗粒特异性感染干细胞的方法，该方法可以递送用于活细胞成像的视觉标志物以及用于富集iPS细胞的选择性标志物。识别SSEA4和CD24的抗体介导iPS细胞对亲本人成纤维细胞的选择性感染，从而允许通过嘌呤霉素选择快速扩增这些细胞。载体的适应允许选择性标记人胚胎干（hES）细胞以将其从分化细胞群中去除。该方法的优点在于，它不仅鉴定干细胞，而且可以将特定基因（包括阳性和阴性选择标记物、调节基因或miRNA）递送至靶向干细胞。特异性靶向基因递送至人多能干细胞的能力在组织工程和干细胞疗法中具有广泛的应用。
The identification of stem cells within a mixed population of cells is a major hurdle for stem cell biology–in particular, in the identification of induced pluripotent stem (iPS) cells during the reprogramming process. Based on the selective expression of stem cell surface markers, a method to specifically infect stem cells through antibody-conjugated lentiviral particles has been developed that can deliver both visual markers for live-cell imaging as well as selectable markers to enrich for iPS cells. Antibodies recognizing SSEA4 and CD24 mediated the selective infection of the iPS cells over the parental human fibroblasts, allowing for rapid expansion of these cells by puromycin selection. Adaptation of the vector allows for the selective marking of human embryonic stem (hES) cells for their removal from a population of differentiated cells. This method has the benefit that it not only identifies stem cells, but that specific genes, including positive and negative selection markers, regulatory genes or miRNA can be delivered to the targeted stem cells. The ability to specifically target gene delivery to human pluripotent stem cells has broad applications in tissue engineering and stem cell therapies.
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