Multiplexed Fluid Flow Device to Study Cellular Response to Tunable Shear Stress Gradients.
Multiplexed Fluid Flow Device to Study Cellular Response to Tunable Shear Stress Gradients.
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DOI：
10.1007/s10439-015-1500-7
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发表时间：
2016-07
影响因子：
3.8
通讯作者：
Dunn AR
中科院分区：
文献类型：
--
作者：
Ostrowski MA;Huang EY;Surya VN;Poplawski C;Barakat JM;Lin GL;Fuller GG;Dunn AR
内皮细胞 (EC) 排列在血管和淋巴管的内部，并经历空间变化的壁剪切应力 (WSS)，这是其生理功能的固有部分。 EC 和哺乳动物细胞一般如何感知空间变化的 WSS 仍然知之甚少，部分原因是缺乏将细胞暴露于空间变化的流动模式的便利工具。我们构建了一个多重设备，称为 6 孔冲击流动室，它向六孔组织培养板提供受控的 WSS 梯度。使用该装置，我们研究了淋巴微血管 EC、脐静脉 EC、原代成纤维细胞和上皮细胞在数小时至数天的时间尺度上对 WSS 梯度的迁移反应。我们观察到淋巴微血管内皮细胞逆着流动方向向上游迁移，这种反应在本文研究的所有细胞类型中是独特的。延时活细胞成像显示，微管组织中心响应所施加的 WSS 梯度而重新定位到细胞核的上游侧。为了进一步证明我们的设备的实用性，我们筛选了规范信号通路在介导这种上游迁移反应中的参与。这些数据强调了 WSS 幅度和 WSS 空间梯度在指示细胞对流体流动的反应中的重要性。
Endothelial cells (ECs) line the interior of blood and lymphatic vessels and experience spatially varying wall shear stress (WSS) as an intrinsic part of their physiological function. How ECs, and mammalian cells generally, sense spatially varying WSS remains poorly understood, due in part to a lack of convenient tools for exposing cells to spatially varying flow patterns. We built a multiplexed device, termed a 6-well impinging flow chamber, that imparts controlled WSS gradients to a six-well tissue culture plate. Using this device, we investigated the migratory response of lymphatic microvascular ECs, umbilical vein ECs, primary fibroblasts, and epithelial cells to WSS gradients on hours to days timescales. We observed that lymphatic microvascular ECs migrate upstream, against the direction of flow, a response that was unique among all the cells types investigated here. Time-lapse, live cell imaging revealed that the microtubule organizing center relocated to the upstream side of the nucleus in response to the applied WSS gradient. To further demonstrate the utility of our device, we screened for the involvement of canonical signaling pathways in mediating this upstream migratory response. These data highlight the importance of WSS magnitude and WSS spatial gradients in dictating the cellular response to fluid flow.