Microfluidic investigation of BDNF-enhanced neural stem cell chemotaxis in CXCL12 gradients.
Microfluidic investigation of BDNF-enhanced neural stem cell chemotaxis in CXCL12 gradients.
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在 CXCL12 梯度中对 BDNF 增强神经干细胞趋化性的微流控研究
DOI：
10.1002/smll.201202208
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发表时间：
2013-02-25
期刊：
影响因子：
13.3
通讯作者：
Heilshorn, Sarah C.
中科院分区：
文献类型：
--
作者：
Xu, Hui;Heilshorn, Sarah C.
体内研究表明，CXCL 12（又名基质细胞衍生因子1α）的梯度可能是脑发育和神经组织再生过程中神经干细胞（NSC）迁移的关键指导线索。然而，传统的体外趋化工具通常受到不稳定的浓度梯度以及无法分离细胞迁移方向和速度的限制。这些局限性限制了神经元迁移的可重复性和定量分析，这是基于机制的研究的要求，可以指导神经再生的新治疗策略的开发。使用微流控梯度发生器，我们定量巢蛋白和Sox-2阳性人胚胎NSC趋化性内的线性和稳定的CXCL 12梯度。虽然未处理的NSC不能在CXCL 12梯度内趋化，但用脑源性神经营养因子（BDNF）预处理细胞导致显著的趋化性、定向迁移。BDNF预处理对细胞迁移速度没有影响，平均迁移速度约为1 μm min-1。定量分析确定CXCL 12浓度高于72 ng ml−1（9.0 nM）高于最小激活阈值，而浓度低于117 ng ml−1（14.7 nM）低于饱和阈值。有趣的是，尽管AMD 3100的抑制剂研究显示CXCL 12趋化性需要受体CXCR 4活化，但发现BDNF预处理对CXCR 4或推定的CXCR 7清道夫受体的mRNA水平或表面呈递没有显著影响。在稳定的趋化因子浓度曲线内NSC迁移的微流体研究提供了定量分析以及对BDNF诱导的趋化性对CXCL 12的迁移机制的新见解。这些数据将为临床组织再生治疗提供定量指导，以制定新的策略来增强内源性和外源性神经干细胞的归巢。
In vivo studies have suggested that gradients of CXCL12 (aka stromal cell-derived factor 1α) may be a critical guidance cue for neural stem cell (NSC) migration during both brain development and neural tissue regeneration. However, traditional in vitro chemotaxis tools are typically limited by unstable concentration gradients and the inability to decouple cell migration directionality and speed. These limitations have restricted the reproducible and quantitative analysis of neuronal migration, which is a requirement for mechanism-based studies that may guide the development of new therapeutic strategies for neural regeneration. Using a microfluidic gradient generator, we quantified nestin and Sox-2 positive human embryonic NSC chemotaxis within a linear and stable CXCL12 gradient. While untreated NSCs were not able to chemotax within CXCL12 gradients, pre-treatment of the cells with brain-derived neurotrophic factor (BDNF) resulted in significant chemotactic, directional migration. BDNF pre-treatment had no effect on cell migration speed, which averaged about 1 μm min−1. Quantitative analysis determined that CXCL12 concentrations above 72 ng ml−1 (9.0 nM) are above the minimum activation threshold, while concentrations below 117 ng ml−1 (14.7 nM) are below the saturation threshold. Interestingly, although inhibitor studies with AMD 3100 revealed that CXCL12 chemotaxis requires receptor CXCR4 activation, BDNF pre-treatment was found to have no profound effects on the mRNA levels or surface presentation of CXCR4 or the putative CXCR7 scavenger receptor. The microfluidic study of NSC migration within stable chemokine concentration profiles provided quantitative analysis as well as new insight into the migratory mechanism underlying BDNF-induced chemotaxis towards CXCL12. These data will provide quantitative guidance in the development of new strategies to enhance the homing of endogenous and exogenous NSCs for clinical tissue regeneration therapies.
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DOI：
10.1634/stemcells.2008-0519
发表时间：
2008-12
期刊：
Stem cells (Dayton, Ohio)
影响因子：
--
作者：
Barkho BZ;Munoz AE;Li X;Li L;Cunningham LA;Zhao X
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DOI：
10.1039/c0ib00038h
发表时间：
2010-11
期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
--
作者：
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通讯作者：
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影响因子：
8.3
作者：
Andres RH;Choi R;Pendharkar AV;Gaeta X;Wang N;Nathan JK;Chua JY;Lee SW;Palmer TD;Steinberg GK;Guzman R
通讯作者：
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影响因子：
5.3
作者：
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通讯作者：
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影响因子：
6.1
作者：
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