The dark side of BrdU in neural stem cell biology: detrimental effects on cell cycle, differentiation and survival
The dark side of BrdU in neural stem cell biology: detrimental effects on cell cycle, differentiation and survival
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DOI：
10.1007/s00441-011-1213-7
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发表时间：
2011-09-01
影响因子：
3.6
通讯作者：
Aigner, Ludwig
中科院分区：
文献类型：
--
作者：
Lehner, Bernadette;Sandner, Beatrice;Aigner, Ludwig
5-溴-2‘-脱氧尿苷(BrdU)广泛应用于神经干细胞生物学分析，尤其是对分裂细胞进行标记和定位。然而，到目前为止，只有少数研究解决了BrdU标记本身是否会影响待研究细胞的问题。在这里，我们重点研究BrdU对成年大鼠脑神经细胞培养和体内祖细胞增殖的潜在影响。体外用BrdU冲击神经球48h，分析细胞增殖、细胞周期、分化、存活和黏附特性。MTS法检测BrdU抑制神经前体细胞的增殖，并增加细胞周期中G0/G1期细胞的比例。此外，BrdU增加了细胞死亡，并以剂量依赖的方式诱导了神经干细胞的黏附。细胞黏附伴随着活性基质金属蛋白酶-2(MMP2)的减少。此外，BrdU抑制神经元和少突胶质细胞的分化，而星形胶质细胞的命运不受影响。与体外情况相比，BrdU显然不会影响体内神经前体细胞的内源性增殖或神经发生，这种浓度通常用于标记神经前体细胞。我们的结果揭示了到目前为止BrdU对成人鼻咽癌细胞的未知作用。我们的结论是，由于BrdU在干细胞生物学中的普遍使用，在解释数据之前，必须仔细检查NPC的BrdU的任何潜在影响。
5-Bromo-2'-deoxyuridin (BrdU) is frequently used in anaylsis of neural stem cell biology, in particular to label and to fate-map dividing cells. However, up to now, only a few studies have addressed the question as to whether BrdU labeling per se affects the cells to be investigated. Here, we focused on the potential impact of BrdU on neurosphere cultures derived from the adult rat brain and on proliferation of progenitors in vivo. In vitro, neurospheres were pulsed for 48 h with BrdU, and cell proliferation, cell cycle, differentiation, survival and adhesion properties were subsequently analyzed. BrdU inhibited the expansion of neural progenitors as assessed by MTS assay and increased the fraction of cells in the G0/G1-phase of the cell cycle. Moreover, BrdU increased cell death and dose-dependently induced adherence of NPCs. Cell adherence was accompanied by a reduced amount of active matrix-metalloproteinase-2 (MMP-2). Furthermore, BrdU repressed neuronal and oligodendroglial differentiation, whereas astroglial fate was not affected. In contrast to the in vitro situation, BrdU apparently did not influence endogenous proliferation of NPCs or neurogenesis in concentrations that are typically used for labeling of neural progenitors in vivo. Our results reveal so far uncharacterized effects of BrdU on adult NPCs. We conclude that, because of its ubiquitous use in stem cell biology, any potential effect of BrdU of NPCs has to be scrutinized prior to interpretation of data.