Spontaneous fusion and nonclonal growth of adult neural stem cells
Spontaneous fusion and nonclonal growth of adult neural stem cells
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DOI：
10.1634/stemcells.2006-0620
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发表时间：
2007-01-01
期刊：
影响因子：
5.2
通讯作者：
Gage, Fred H.
中科院分区：
文献类型：
--
作者：
Jessberger, Sebastian;Clemenson, Gregory D., Jr.;Gage, Fred H.
多能神经干细胞（NSC）可以从成人脑的各个区域分离并在体外增殖。最近的报告表明，自发的融合事件之间的神经干细胞生长时，作为自由浮动的神经球，可能会影响神经干细胞文化的遗传组成。我们使用表达红色荧光蛋白（RFP）或绿色荧光蛋白（GFP）的成体神经干细胞来分析大鼠和小鼠神经干细胞的融合频率。荧光激活细胞分选（FACS）显示，在增殖条件下，约0.2%的大鼠和小鼠神经干细胞共表达RFP和GFP，无论细胞是否生长为神经球（小鼠神经干细胞）或附着单层（大鼠和小鼠神经干细胞）。融合的细胞不增殖，不能繁殖，表明异常融合的细胞是没有活力的。此外，我们发现，无论是神经球，也不是单层克隆生长，因为即使是非常低密度的文化有球体含有GFP和RFP表达细胞和单层补丁与GFP和RFP表达细胞在密切的接近。不同NSC群体之间的非克隆生长强烈表明使用仔细和精确的培养条件，如单细胞测定，以表征体外NSC的效力和生长。
Multipotent neural stem cells (NSCs) can be isolated from various regions of the adult brain and propagated in vitro. Recent reports have suggested spontaneous fusion events among NSCs when grown as free-floating neurospheres that may affect the genetic composition of NSC cultures. We used adult NSCs expressing either red fluorescent protein (RFP) or green fluorescent protein (GFP) to analyze the fusion frequency of rat and mouse NSCs. Fluorescence-activated cell sorting (FACS) revealed that, under proliferating conditions, approximately 0.2% of rat and mouse NSCs coexpressed RFP and GFP irrespective of whether the cells were grown as neurospheres (mouse NSCs) or as attached monolayers (rat and mouse NSCs). Fused cells did not proliferate and could not be propagated, suggesting that aberrantly fused cells are not viable. Furthermore, we found that neither neurospheres nor monolayers grew clonally, because even very low-density cultures had spheres containing both GFP- and RFP-expressing cells and monolayer patches with GFP- and RFP-expressing cells in close proximity. The nonclonal growth between distinct NSC populations strongly suggests the use of careful and precise culture conditions, such as single-cell assays, to characterize potency and growth of NSCs in vitro.