Cell density-dependent differential proliferation of neural stem cells on omnidirectional nanopore-arrayed surface.
Cell density-dependent differential proliferation of neural stem cells on omnidirectional nanopore-arrayed surface.
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DOI：
10.1038/s41598-017-13372-6
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发表时间：
2017-10-12
影响因子：
4.6
通讯作者：
Kim HJ
中科院分区：
文献类型：
--
作者：
Cha KJ;Kong SY;Lee JS;Kim HW;Shin JY;La M;Han BW;Kim DS;Kim HJ
最近，表面纳米拓扑学在决定干细胞命运和行为方面的重要性已经被揭示出来。在本研究中，我们制作了具有全方位纳米孔阵列表面的聚苯乙烯细胞培养皿(直径：200 nm，深度：500 nm，中心到中心距离：500 nm)，并研究了纳米拓扑术对大鼠神经干细胞(NSCs)的影响。当细胞密度较低时，神经干细胞在ONAS上的增殖能力高于在平坦表面上培养的NSCs，在有丝分裂原存在的条件下，NSCs在增殖过程中的自发分化程度较低。有趣的是，在ONAS上以克隆密度培养的NSCs显示出生成神经球的倾向，而那些在平面上的NSCs迁移出去，作为个体增殖，并扩散到表面附着。然而，不同的增殖模式是依赖于细胞密度的，因为随着细胞密度的增加，明显的现象消失了。ONAS调节细胞骨架重组并抑制焦点粘连的形成，这在生长在平面上的神经干细胞中普遍观察到。ONAS似乎加强了NSC-NSC的相互作用，限制了单个细胞的迁移，并抑制了NSC在纳米孔表面的附着。这些数据表明，ONAS保持了NSCs的未分化状态，同时保持了多能性，是培养低密度NSCs的较好地形。
Recently, the importance of surface nanotopography in the determination of stem cell fate and behavior has been revealed. In the current study, we generated polystyrene cell-culture dishes with an omnidirectional nanopore arrayed surface (ONAS) (diameter: 200 nm, depth: 500 nm, center-to-center distance: 500 nm) and investigated the effects of nanotopography on rat neural stem cells (NSCs). NSCs cultured on ONAS proliferated better than those on the flat surface when cell density was low and showed less spontaneous differentiation during proliferation in the presence of mitogens. Interestingly, NSCs cultured on ONAS at clonal density demonstrated a propensity to generate neurospheres, whereas those on the flat surface migrated out, proliferated as individuals, and spread out to attach to the surface. However, the differential patterns of proliferation were cell density-dependent since the distinct phenomena were lost when cell density was increased. ONAS modulated cytoskeletal reorganization and inhibited formation of focal adhesion, which is generally observed in NSCs grown on flat surfaces. ONAS appeared to reinforce NSC-NSC interaction, restricted individual cell migration and prohibited NSC attachment to the nanopore surface. These data demonstrate that ONAS maintains NSCs as undifferentiated while retaining multipotency and is a better topography for culturing low density NSCs.
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