A human iPSC line capable of differentiating into functional macrophages expressing ZsGreen: a tool for the study and in vivo tracking of therapeutic cells.
A human iPSC line capable of differentiating into functional macrophages expressing ZsGreen: a tool for the study and in vivo tracking of therapeutic cells.
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DOI：
10.1098/rstb.2017.0219
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发表时间：
2018-07-05
期刊：
影响因子：
--
通讯作者：
Forrester LM
中科院分区：
文献类型：
--
作者：
Lopez-Yrigoyen M;Fidanza A;Cassetta L;Axton RA;Taylor AH;Meseguer-Ripolles J;Tsakiridis A;Wilson V;Hay DC;Pollard JW;Forrester LM
我们描述了一种人类诱导多能干细胞(IPSC)系SFCi55-ZsGr的产生，该系被设计成以结构性方式表达荧光报告基因ZsGreen。将CAG驱动的ZsGreen表达盒插入AAVS1基因座，并在未分化的IPSCs和来自所有三个生殖层的细胞系(包括造血细胞、肝细胞和神经元)中观察到高水平的表达。我们证明了从SFCi55-ZsGreen iPSC系产生的终末分化巨噬细胞的有效产生，并表明它们在基因表达、细胞表面标记表达和吞噬活性方面与其亲本SFCi55 iPSC系产生的巨噬细胞没有区别。ZsGreen的高水平表达对巨噬细胞对M(脂多糖+干扰素γ)、M(IL10)或M(IL4)表型的激活能力没有影响，也没有影响其可塑性，通过评估它们从一种表型转换到另一种表型的能力。因此，通过靶向IPSCs中的AAVS1基因座，可以生产出功能齐全、荧光标记的人巨噬细胞，用于疾病模型的体内追踪。该战略还提供了一个平台，用于引入预计将调节和/或稳定巨噬细胞功能的因子。本文是主题期刊《人体组织设计者：来到你身边的实验室》的一部分。
We describe the production of a human induced pluripotent stem cell (iPSC) line, SFCi55-ZsGr, that has been engineered to express the fluorescent reporter gene, ZsGreen, in a constitutive manner. The CAG-driven ZsGreen expression cassette was inserted into the AAVS1 locus and a high level of expression was observed in undifferentiated iPSCs and in cell lineages derived from all three germ layers including haematopoietic cells, hepatocytes and neurons. We demonstrate efficient production of terminally differentiated macrophages from the SFCi55-ZsGreen iPSC line and show that they are indistinguishable from those generated from their parental SFCi55 iPSC line in terms of gene expression, cell surface marker expression and phagocytic activity. The high level of ZsGreen expression had no effect on the ability of macrophages to be activated to an M(LPS + IFNγ), M(IL10) or M(IL4) phenotype nor on their plasticity, assessed by their ability to switch from one phenotype to another. Thus, targeting of the AAVS1 locus in iPSCs allows for the production of fully functional, fluorescently tagged human macrophages that can be used for in vivo tracking in disease models. The strategy also provides a platform for the introduction of factors that are predicted to modulate and/or stabilize macrophage function. This article is part of the theme issue ‘Designer human tissue: coming to a lab near you’.
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影响因子：
5.9
作者：
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5.9
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