A microfluidic platform for generation of sharp gradients in open-access culture
A microfluidic platform for generation of sharp gradients in open-access culture
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DOI：
10.1063/1.3490784
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发表时间：
2010-12-01
期刊：
影响因子：
3.2
通讯作者：
Folch, Albert
中科院分区：
文献类型：
--
作者：
Cate, David M.;Sip, Christopher G.;Folch, Albert
控制培养细胞的3D微环境对于理解生物分子浓度梯度对细胞生长、再生和分化的复杂关系至关重要。本文报道了一种微流控装置，用于将可溶性分子的梯度递送到开放储液器中的细胞，而不使细胞暴露于流动。细胞在聚酯膜上培养，该聚酯膜将细胞与提供梯度的流动隔离。实现了一种新颖的“盖”设计，其防止膜周围的泄漏，而不需要密封剂或粘合剂。一旦层被模制，器件制造可以在室温下在几分钟内进行。表面梯度，其特征在于与落射荧光显微镜;图像分析证实，尖锐的梯度（类似于33 μ m宽）可以重现产生。我们表明，应用于膜下的橙子和绿色细胞跟踪器（CT）的非均质层流模式可以定位于另一侧培养的细胞;浓度分布扫描显示CT扩散平行于膜表面的程度为10-20 μ m。我们的装置对于常规细胞培养是理想的，因为细胞培养表面易于物理操作（例如，微量移液管进入），细胞培养基与培养箱气氛直接接触（即，不需要用于确保气体浓度适当平衡的特殊方案），并且细胞不经受流动诱导的剪切力，这是在封闭通道微流体设计中不常见的有利属性。(C)2010年美国物理学会。[doi 10.1063/1.3490784]
Control of the 3D microenvironment for cultured cells is essential for understanding the complex relationships that biomolecular concentration gradients have on cellular growth, regeneration, and differentiation. This paper reports a microfluidic device for delivering gradients of soluble molecules to cells in an open reservoir without exposing the cells to flow. The cells are cultured on a polyester membrane that shields them from the flow that delivers the gradient. A novel "lid" design is implemented which prevents leakage from around the membrane without requiring sealing agents or adhesives. Once layers are molded, device fabrication can be performed within minutes while at room temperature. Surface gradients were characterized with epifluorescence microscopy; image analysis verified that sharp gradients (similar to 33 mu m wide) can be reproducibly generated. We show that heterogeneous laminar flow patterns of Orange and Green Cell Tracker (CT) applied beneath the membrane can be localized to cells cultured on the other side; concentration profile scans show the extent of CT diffusion parallel to the membrane's surface to be 10-20 mu m. Our device is ideal for conventional cell culture because the cell culture surface is readily accessible to physical manipulation (e.g., micropipette access), the cell culture medium is in direct contact with the incubator atmosphere (i.e., no special protocols for ensuring proper equilibration of gas concentrations are required), and the cells are not subjected to flow-induced shear forces, which are advantageous attributes not commonly found in closed-channel microfluidic designs. (C) 2010 American Institute of Physics. [doi:10.1063/1.3490784]