Isolation and characterization of mammalian cells expressing the Arf promoter during eye development.
Isolation and characterization of mammalian cells expressing the Arf promoter during eye development.
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DOI：
10.2144/000114166
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发表时间：
2014-05
期刊：
影响因子：
2.7
通讯作者：
Skapek SX
中科院分区：
文献类型：
--
作者：
Iqbal NS;Xu L;Devitt CC;Skapek SX
虽然许多研究人员利用各种类型的培养的癌细胞和永生化的成纤维细胞成功地发现了肿瘤抑制基因p19Arf的新功能，但这些系统并不能准确地反映Arf在发育过程中表达的内源环境。我们通过从小鼠眼睛的初级玻璃体中分离血管周围细胞来解决这个问题。这些细胞代表了一种罕见的细胞类型，通常在非病理的发育环境中表达p19Arf肿瘤抑制因子。我们利用荧光激活的细胞分选法，通过天然Arf启动子驱动的GFP报告基因来纯化细胞，并对其形态和基因表达模式进行了表征。我们进一步研究了Arf在PVCs中重新引入的效果，以验证预期的p19Arf下游效应器，并揭示作为血管生成调节的新功能。这种方法学和细胞培养模型可以作为研究p19Arf生物学的有用工具。
Although many researchers have successfully uncovered novel functions of the tumor suppressor p19Arf utilizing various types of cultured cancer cells and immortalized fibroblasts, these systems do not accurately reflect the endogenous environment in which Arf is developmentally expressed. We addressed this by isolating perivascular cells from the primary vitreous of the mouse eye. These cells represent a rare cell type that normally expresses the p19Arf tumor suppressor in a non-pathological, developmental context. We utilized fluorescence activated cell sorting to purify the cells by virtue of a GFP reporter driven by the native Arf promoter, and characterized their morphology and gene expression pattern. We further examined the effects of reintroduction of Arf in the PVCs to verify expected downstream effectors of p19Arf as well as uncover novel functions as a regulator of vasculogenesis. This methodology and cell culture model should serve as a useful tool to examine p19Arf biology.
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影响因子：
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作者：
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影响因子：
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