Interactions between multiple cell types in parallel microfluidic channels: Monitoring platelet adhesion to an endothelium in the presence of an anti-adhesion drug
Interactions between multiple cell types in parallel microfluidic channels: Monitoring platelet adhesion to an endothelium in the presence of an anti-adhesion drug
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DOI：
10.1021/ac801114j
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发表时间：
2008-10-01
影响因子：
7.4
通讯作者：
Spence, Dana M.
中科院分区：
文献类型：
--
作者：
Ku, Chia-Jui;Oblak, Teresa D'Amico;Spence, Dana M.
提出了一种用软光刻技术在微流控装置的通道中固定内皮细胞的简单方法，该方法不需要对与微流控装置结合使用的衬底材料进行表面氧化，并且即使使用可逆密封也可以操作。研究了牛肺动脉内皮细胞（bPAECs）在培养皿表面的最佳培养条件。研究的参数包括纤维连接蛋白浓度、温度、播种密度和固定时间。为了提高设备的实用性，所有的优化研究，以及涉及血小板粘附到固定内皮的研究，都是在平行通道中进行的，从而提高了单通道设备的吞吐量。细胞固定的最佳条件为：在培养皿上涂覆100 μ g/mL的纤维连接蛋白，播种细胞密度为1.00 × 10(5)个细胞mL(-1)，固定时间为90 min，温度为37℃。然后在已知的血小板激活剂（ADP）和血小板活化药物抑制剂存在的情况下，利用该装置监测血小板与固定内皮的物理相互作用（粘附）。通道内粘附内皮细胞的血小板数量从无ADP时的17.0 +/- 2.3增加到有5.00 μ M ADP时的63.2 +/- 2.4。此外，本文的数据还显示，抑制内皮细胞一氧化氮（NO）的产生（一种公认的血小板粘附内皮的抑制剂）可使粘附在内皮表面的血小板数量增加到35.4 +/- 1.0。在NO抑制和5.00 μ M ADP存在的情况下，对血小板粘附的影响进一步增加到127 +/- 5.2。最后，利用该装置研究了一种已知的抑制血小板粘附的药物（氯吡格雷）的作用，在药物存在的情况下，由于5.00 μ M ADP激活的血小板粘附降低到24.0 +/- 3.8。这项工作首次展示了多种细胞类型在微流体装置的通道中物理相互作用，并进一步证明了这些装置在药物发现过程和药物功效研究中的潜力。
A simple method for immobilizing endothelial cells in the channels of a microfluidic device fabricated with soft lithography is presented that requires no surface oxidation of the substrate material used in conjunction with the microfluidic device and is operable even with a reversible seal. Specifically, optimal conditions for culturing bovine pulmonary artery endothelial cells (bPAECs) to the surface of a Petri dish were investigated. The parameters investigated included fibronectin concentration, temperature, seeding density, and immobilization time. To enhance the utility of the device, all optimization studies, and studies involving platelet adhesion to the immobilized endothelium, were performed in parallel channels, thereby enabling improved throughput over a single channel device. The optimal conditions for cell immobilization included coating the Petri dish with 100 mu g/mL fibronectin, a seeding cell density of 1.00 x 10(5) cells mL(-1), and an immobilization time of 90 min at 37 degrees C. The device was then employed to monitor the physical interaction (adhesion) of platelets to the immobilized endothelium in the presence of a known platelet activator (ADP) and a drug inhibitor of platelet activation. The number of platelets adhering to the endothelial cells in the channels increased from 17.0 +/- 2.3 in the absence of ADP to 63.2 +/- 2.4 in the presence of 5.00 mu M ADP. Moreover, the data presented here also shows that inhibition of endothelium nitric oxide (NO) production, a recognized inhibitor of platelet adhesion to the endothelium, increased the number of platelets adhering to the surface to 35.4 +/- 1.0. In the presence of NO inhibition and 5.00 mu M ADP, the affect on platelet adhesion was further increased to 127 +/- 5.2. Finally, this device was employed to investigate the effect of a drug known to inhibit platelet adhesion (clopidogrel) and, in the presence of the drug, the platelet adhesion due to activation by 5.00 mu M ADP decreased to 24.0 +/- 3.8. This work is the first representation of multiple cell types physically interacting in the channels of a microfluidic device and further demonstrates the potential of these devices in the drug discovery process and drug efficacy studies.