Biomicrofluidics. 2011 Jun;5(2):22214. doi: 10.1063/1.3580756. Epub 2011 Jun 29.
In this study, we developed a miniaturized microfluidic-based high-throughput cell toxicity assay to create an in vitro model of Parkinson's disease (PD). In particular, we generated concentration gradients of 6-hydroxydopamine (6-OHDA) to trigger a process of neuronal apoptosis in pheochromocytoma PC12 neuronal cell line. PC12 cells were cultured in a microfluidic channel, and a concentration gradient of 6-OHDA was generated in the channel by using a back and forth movement of the fluid flow. Cellular apoptosis was then analyzed along the channel. The results indicate that at low concentrations of 6-OHDA along the gradient (i.e., approximately less than 260 μM), the neuronal death in the channel was mainly induced by apoptosis, while at higher concentrations, 6-OHDA induced neuronal death mainly through necrosis. Thus, this concentration appears to be useful for creating an in vitro model of PD by inducing the highest level of apoptosis in PC12 cells. As microfluidic systems are advantageous in a range of properties such as throughput and lower use of reagents, they may provide a useful approach for generating in vitro models of disease for drug discovery applications.
在这项研究中，我们开发了一种基于微流控的小型高通量细胞毒性测定法，以建立帕金森病（PD）的体外模型。具体来说，我们生成了 6-羟多巴胺（6-OHDA）的浓度梯度，以触发嗜铬细胞瘤 PC12 神经元细胞系的神经元凋亡过程。PC12 细胞在微流控通道中培养，并通过流体的来回运动在通道中产生 6-OHDA 的浓度梯度。然后沿通道分析细胞凋亡。结果表明，在梯度中较低浓度的 6-OHDA（即约小于 260μM）下，通道中的神经元死亡主要是由凋亡引起的，而在较高浓度下，6-OHDA 主要通过坏死诱导神经元死亡。因此，这个浓度似乎可以通过诱导 PC12 细胞中最高水平的凋亡来创建 PD 的体外模型。由于微流控系统在通量和试剂使用量等方面具有优势，因此它们可能为药物发现应用提供生成疾病体外模型的有用方法。
