Mouthon Marc-André, Fouchet Pierre, Mathieu Céline, Sii-Felice Karine, Etienne Olivier, Lages Céline Silva, Boussin François D
Laboratoire de RadioPathologie CEA/DSV/DRR-IPSC, Fontenay-aux-Roses cedex, France.
J Neurochem. 2006 Nov;99(3):807-17. doi: 10.1111/j.1471-4159.2006.04118.x. Epub 2006 Aug 21.
Developing and adult forebrains contain neural stem cells (NSCs) but no marker is available to highly purify them. When analysed by flow cytometry, stem cells from various tissues are enriched in a 'side population' (SP) characterized by the exclusion of the fluorescent dye Hoechst 33342. Here, we characterize the SP in embryonic, neonatal and adult forebrains, as well as in neurosphere cultures and we have determined whether this SP could be a source of enriched NSCs. By using specific inhibitors, we found that the SP from embryonic forebrain results from the activity of the ABCG2 transporter, a characteristic of other stem cells, whereas the SP from adult forebrain probably results from the ABCB1 transporter. SP cells from embryonic and adult forebrains, however, expressed a range of cell surface markers more consistent with a haematopoietic/endothelial origin than with a neural origin; NSC markers were mostly expressed on cells outside the SP (in the main population, MP). Moreover, assays for NSC growth in vitro showed that SP cells from embryonic and adult forebrains did not generate NSC-derived colonies, whereas the MP did. We thus conclude that NSCs from developing and adult forebrains are not contained in the SP contrary to stem cells from other tissues.
发育中和成年的前脑含有神经干细胞（NSCs），但目前尚无用于高度纯化这些细胞的标志物。当通过流式细胞术分析时，来自各种组织的干细胞在一个“侧群”（SP）中富集，该侧群的特征是能够排斥荧光染料Hoechst 33342。在这里，我们对胚胎、新生和成年前脑以及神经球培养物中的SP进行了表征，并确定了这个SP是否可能是富集神经干细胞的来源。通过使用特异性抑制剂，我们发现胚胎前脑的SP是由ABCG2转运蛋白的活性产生的，这是其他干细胞的一个特征，而成年前脑的SP可能是由ABCB1转运蛋白产生的。然而，胚胎和成年前脑的SP细胞表达了一系列细胞表面标志物，这些标志物与造血/内皮起源更一致，而不是与神经起源一致；神经干细胞标志物大多表达在SP之外的细胞上（在主要群体，即MP中）。此外，体外神经干细胞生长测定表明，胚胎和成年前脑的SP细胞不会产生神经干细胞衍生的集落，而MP细胞则会。因此，我们得出结论，与其他组织的干细胞不同，发育中和成年前脑的神经干细胞并不存在于SP中。
