A novel fluorescent reporter CDy1 enriches for neural stem cells derived from the murine brain.
A novel fluorescent reporter CDy1 enriches for neural stem cells derived from the murine brain.
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一种新型荧光报告基因 CDy1 富集源自小鼠大脑的神经干细胞。
DOI：
10.1089/scd.2012.0660
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发表时间：
2013
影响因子：
4
通讯作者：
G. Osborne
中科院分区：
文献类型：
--
作者：
J. Vukovic;A. Bedin;P. Bartlett;G. Osborne
成年哺乳动物的两个脑区持续发生神经发生，并有大量可分化为所有神经细胞类型的神经干细胞（NSCs）作为支撑。为了进一步了解NSC的功能，开发新的治疗和诊断方法，准确地鉴定和富集NSCs是非常重要的。目前还没有明确的标记物来鉴定和富集存在于小鼠大脑中的NSCs。最近，一种荧光迷迭胺染料CDy1已被确定为培养的人类胚胎和诱导多能干细胞的多能性标记。由于类似的细胞特性可能使CDy1被其他干细胞群摄取和保留，我们假设这种染料也可能富集于小鼠大脑的原代NSCs。因为在成年哺乳动物中，脑室下区（SVZ）和海马体代表了NSCs高度富集的大脑区域，我们从这些区域取样细胞来验证这一假设。这些实验表明，CDy1染色确实允许从SVZ和海马中富集和选择所有神经球形成细胞。接下来，我们检测了CDy1在选择老年动物SVZ衍生的NSCs方面的有效性，老年动物SVZ中存在的NSCs总数明显低于年轻动物。我们发现CDy1有效地标记成年动物和老年动物的NSCs，通过神经球测定，并反映老年动物中存在的NSCs数量。因此，CDy1似乎是原发性脑组织制备中NSCs富集的新标记物。
Neurogenesis occurs continuously in two brain regions of adult mammals, underpinned by a pool of resident neural stem cells (NSCs) that can differentiate into all neural cell types. To advance our understanding of NSC function and to develop therapeutic and diagnostic approaches, it is important to accurately identify and enrich for NSCs. There are no definitive markers for the identification and enrichment of NSCs present in the mouse brain. Recently, a fluorescent rosamine dye, CDy1, has been identified as a label for pluripotency in cultured human embryonic and induced pluripotent stem cells. As similar cellular characteristics may enable the uptake and retention of CDy1 by other stem cell populations, we hypothesized that this dye may also enrich for primary NSCs from the mouse brain. Because the subventricular zone (SVZ) and the hippocampus represent brain regions that are highly enriched for NSCs in adult mammals, we sampled cells from these areas to test this hypothesis. These experiments revealed that CDy1 staining indeed allows for enrichment and selection of all neurosphere-forming cells from both the SVZ and the hippocampus. We next examined the effectiveness of CDy1 to select for NSCs derived from the SVZ of aged animals, where the total pool of NSCs present is significantly lower than in young animals. We found that CDy1 effectively labels the NSCs in adult and aged animals as assessed by the neurosphere assay and reflects the numbers of NSCs present in aged animals. CDy1, therefore, appears to be a novel marker for enrichment of NSCs in primary brain tissue preparations.
影响因子：
14.5
作者：
Deleyrolle, Loic P.;Harding, Angus;Reynolds, Brent A.
通讯作者：
Reynolds, Brent A.
影响因子：
2.7
作者：
Capela, A;Temple, S
通讯作者：
Temple, S