Microfluidic transwell inserts for generation of tissue culture-friendly gradients in well plates.
Microfluidic transwell inserts for generation of tissue culture-friendly gradients in well plates.
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DOI：
10.1039/c3lc51052b
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发表时间：
2014-01-21
期刊：
影响因子：
6.1
通讯作者：
Folch A
中科院分区：
文献类型：
--
作者：
Sip CG;Bhattacharjee N;Folch A
生化分子的梯度在轴突生长、组织形态发生和跨上皮营养物运输等许多生理过程以及伤口愈合、免疫反应、细菌入侵和癌症转移等病理生理现象中发挥着关键作用。在本文中，我们报告了一种微流体跨孔插入物，用于以用户友好的模块化格式生成可量化的浓度梯度，该格式与传统细胞培养物和组织外植体培养物兼容。该装置只需插入标准 6 孔板中，即可自支撑地悬挂在细胞培养表面上方约 250 μm 的距离处。梯度是由来自设备的小微流通过集成的径迹蚀刻多孔膜进入细胞培养孔而产生的。微流控 Transwell 可以在大面积上以极低的流体剪切应力向在 6 孔板表面独立培养的解离细胞或组织外植体提供稳定、可量化的梯度。我们使用有限元建模来描述多孔膜流动和分子传输，并预测设备产生的梯度。使用该设备，我们将趋化肽 N-Formyl-Met-Leu-Phe (fMLP) 梯度应用于大量 HL-60 细胞（中性粒细胞细胞系），并通过延时显微镜直接观察迁移情况。通过自动跟踪算法对趋化反应进行量化，我们发现 74% 的细胞朝梯度方向移动。此外，模块化设计和低流体剪切应力使得将生长因子和第二信使的梯度应用于小鼠视网膜外植体培养成为可能。微流控 Transwell 装置具有简化的界面和明确的梯度，具有在梯度传感生物学中广泛应用的潜力。
Gradients of biochemical molecules play a key role in many physiological processes such as axon growth, tissue morphogenesis, and trans-epithelium nutrient transport, as well as in pathophysiological phenomena such as wound healing, immune response, bacterial invasion, and cancer metastasis. In this paper, we report a microfluidic transwell insert for generating quantifiable concentration gradients in a user-friendly and modular format that is compatible with conventional cell cultures and with tissue explant cultures. The device is simply inserted into a standard 6-well plate, where it hangs self-supported at a distance of ~250 μm above the cell culture surface. The gradient is created by small microflows from the device, through an integrated track-etched porous membrane, into the cell culture well. The microfluidic transwell can deliver stable, quantifiable gradients over a large area with extremely low fluid shear stress to dissociated cells or tissue explants cultured independently on the surface of a 6-well plate. We used finite-element modeling to describe the porous membrane flow and molecular transport and to predict gradients generated by the device. Using the device, we applied a gradient of the chemotactic peptide N-Formyl-Met-Leu-Phe (fMLP) to a large population of HL-60 cells (a neutrophil cell line) and directly observed the migration with time-lapse microscopy. On quantification of the chemotactic response with an automated tracking algorithm, we found 74% of the cells moving towards the gradient. Additionally, the modular design and low fluid shear stress made it possible to apply gradients of growth factors and second messengers to mouse retinal explant cultures. With a simplified interface and well-defined gradients, the microfluidic transwell device has potential for broad applications to gradient-sensing biology.
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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：
Bhattacharjee N;Li N;Keenan TM;Folch A
通讯作者：
Folch A
影响因子：
2.5
作者：
Ambravaneswaran, Vijayakrishnan;Wong, Ian Y.;Irimia, Daniel
通讯作者：
Irimia, Daniel
影响因子：
7.4
作者：
Bancaud, A;Wagner, G;Viovy, JL
通讯作者：
Viovy, JL
影响因子：
3.2
作者：
Cate, David M.;Sip, Christopher G.;Folch, Albert
通讯作者：
Folch, Albert
DOI：
10.1083/jcb.201010143
发表时间：
2011-05-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Cavnar PJ;Berthier E;Beebe DJ;Huttenlocher A
通讯作者：
Huttenlocher A