Regional and Stage-Specific Effects of Prospectively Purified Vascular Cells on the Adult V-SVZ Neural Stem Cell Lineage
Regional and Stage-Specific Effects of Prospectively Purified Vascular Cells on the Adult V-SVZ Neural Stem Cell Lineage
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DOI：
10.1523/jneurosci.1188-14.2015
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发表时间：
2015-03-18
影响因子：
5.3
通讯作者：
Doetsch, Fiona
中科院分区：
文献类型：
--
作者：
Crouch, Elizabeth E.;Liu, Chang;Doetsch, Fiona
成体神经干细胞存在于专门的小生境中。在脑室-脑室下区（V-SVZ）中，静止神经干细胞（qNSC）被激活（aNSC），并产生转运放大细胞（TAC），其产生迁移到嗅球的成神经细胞。血管系统是成体神经干细胞生态位的重要组成部分，但神经源性区域的血管细胞是否与大脑其他区域的血管细胞本质上不同尚不清楚。此外，周细胞对神经干细胞龛的贡献尚未确定。在这里，我们描述了一种快速的流式细胞仪纯化策略，同时分离的主要内皮细胞和周细胞从非转基因小鼠的脑微区使用CD 31和CD 13作为表面标记。我们比较了纯化的血管细胞从一个神经源性（V-SVZ）和非神经源性脑区（皮质）的V-SVZ干细胞谱系在体外的效果。来自两个区域的内皮和周细胞扩散信号差异性地促进qNSC、aNSC和TAC的增殖和神经元分化。出乎意料的是，可扩散的皮质信号对V-SVZ增殖和神经发生具有最有效的作用，突出了非神经源性脉管系统支持干细胞行为的内在能力。最后，我们确定PlGF-2作为内皮源性促分裂原，促进V-SVZ细胞增殖。这种纯化策略提供了一个平台，以确定在不同的生理和病理状态下血管细胞对干细胞龛和其他脑区域的功能和分子贡献。
Adult neural stem cells reside in specialized niches. In the ventricular-subventricular zone (V-SVZ), quiescent neural stem cells (qNSCs) become activated (aNSCs), and generate transit amplifying cells (TACs), which give rise to neuroblasts that migrate to the olfactory bulb. The vasculature is an important component of the adult neural stem cell niche, but whether vascular cells in neurogenic areas are intrinsically different from those elsewhere in the brain is unknown. Moreover, the contribution of pericytes to the neural stem cell niche has not been defined. Here, we describe a rapid FACS purification strategy to simultaneously isolate primary endothelial cells and pericytes from brain microregions of nontransgenic mice using CD31 and CD13 as surface markers. We compared the effect of purified vascular cells from a neurogenic (V-SVZ) and non-neurogenic brain region (cortex) on the V-SVZ stem cell lineage in vitro. Endothelial and pericyte diffusible signals from both regions differentially promote the proliferation and neuronal differentiation of qNSCs, aNSCs, and TACs. Unexpectedly, diffusible cortical signals had the most potent effects on V-SVZ proliferation and neurogenesis, highlighting the intrinsic capacity of non-neurogenic vasculature to support stem cell behavior. Finally, we identify PlGF-2 as an endothelial-derived mitogen that promotes V-SVZ cell proliferation. This purification strategy provides a platform to define the functional and molecular contribution of vascular cells to stem cell niches and other brain regions under different physiological and pathological states.