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本文引用的文献

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Variation in the safety of induced pluripotent stem cell lines.诱导多能干细胞系安全性的差异。
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Identification of transplantable dopamine neuron precursors at different stages of midbrain neurogenesis.中脑神经发生不同阶段可移植多巴胺能神经元前体的鉴定。
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Efficient production of mesencephalic dopamine neurons by Lmx1a expression in embryonic stem cells.通过胚胎干细胞中Lmx1a的表达高效生成中脑多巴胺能神经元。
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Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling.通过双重抑制SMAD信号通路实现人胚胎干细胞和诱导多能干细胞的高效神经转化。
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Virus-free induction of pluripotency and subsequent excision of reprogramming factors.无病毒诱导多能性及随后重编程因子的切除
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BAC transgenesis in human embryonic stem cells as a novel tool to define the human neural lineage.在人类胚胎干细胞中进行细菌人工染色体转基因作为定义人类神经谱系的一种新工具。
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Striatal progenitors derived from human ES cells mature into DARPP32 neurons in vitro and in quinolinic acid-lesioned rats.源自人类胚胎干细胞的纹状体祖细胞在体外以及喹啉酸损伤的大鼠体内成熟为多巴胺和环磷腺苷调节磷酸蛋白32（DARPP32）神经元。
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Connecting microRNA genes to the core transcriptional regulatory circuitry of embryonic stem cells.将微小RNA基因与胚胎干细胞的核心转录调控回路相连接。
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Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development.胚胎干细胞向临床相关细胞群体的分化：来自胚胎发育的启示
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Identification and targeting of the ROSA26 locus in human embryonic stem cells.人类胚胎干细胞中ROSA26位点的鉴定与靶向作用
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使用 microRNA 调控的慢病毒载体追踪多能培养中的神经前体细胞分化。

Tracking differentiating neural progenitors in pluripotent cultures using microRNA-regulated lentiviral vectors.

机构信息

Department of Experimental Medical Sciences, Wallenberg Neuroscience Center, Lund University, 221 84 Lund, Sweden.

出版信息

Proc Natl Acad Sci U S A. 2010 Jun 22;107(25):11602-7. doi: 10.1073/pnas.1006568107. Epub 2010 Jun 7.

DOI:10.1073/pnas.1006568107
PMID:20534548
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2895063/
Abstract

In this study, we have used a microRNA-regulated lentiviral reporter system to visualize and segregate differentiating neuronal cells in pluripotent cultures. Efficient suppression of transgene expression, specifically in undifferentiated pluripotent cells, was achieved by using a lentiviral vector expressing a fluorescent reporter gene regulated by microRNA-292. Using this strategy, it was possible to track progeny from murine ES, human ES cells, and induced pluripotent stem cells as they differentiated toward the neural lineage. In addition, this strategy was successfully used to FACS purify neuronal progenitors for molecular analysis and transplantation. FACS enrichment reduced tumor formation and increased survival of ES cell-derived neuronal progenitors after transplantation. The properties and versatility of the microRNA-regulated vectors allows broad use of these vectors in stem cell applications.

摘要

在这项研究中，我们使用了一种 microRNA 调控的慢病毒报告系统来可视化和分离多能培养中的分化神经元细胞。通过使用表达受 microRNA-292 调控的荧光报告基因的慢病毒载体，实现了转基因表达的高效抑制，特别是在未分化的多能细胞中。利用这种策略，我们可以追踪从鼠 ES 细胞、人 ES 细胞和诱导多能干细胞分化为神经谱系的后代。此外，该策略还成功地用于 FACS 纯化神经祖细胞进行分子分析和移植。FACS 富集减少了肿瘤形成，并提高了 ES 细胞来源的神经元祖细胞移植后的存活率。microRNA 调控载体的特性和多功能性允许这些载体在干细胞应用中广泛使用。