Microengineering 3D Collagen Hydrogels with Long-Range Fiber Alignment.
Microengineering 3D Collagen Hydrogels with Long-Range Fiber Alignment.
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DOI：
10.3791/64457
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发表时间：
2022-09-07
期刊：
影响因子：
--
通讯作者：
Abhyankar VV
中科院分区：
文献类型：
--
作者：
Ahmed A;Joshi IM;Goulet MR;Vidas JA;Byerley AM;Mansouri M;Day SW;Abhyankar VV
排列的I型胶原纤维引导肿瘤细胞的运动，影响内皮细胞的形态，控制干细胞的分化，是心脏和肌肉骨骼组织的标志。为了研究细胞对体外排列微环境的响应，已经开发了几种方法来产生具有定义的纤维排列的Col1基质，包括磁法、机械法、基于细胞的方法和微流体方法。其中，微流控方法提供了对流体流动和细胞微环境的精确控制等先进能力。然而，为先进的体外培养平台生成排列的CO1基质的微流体方法仅限于延伸距离小于500μm的CO1纤维的薄垫(<；40μm厚)，不利于3D细胞培养应用。在这里，我们提出了一种在微流控器件中制造具有毫米尺度区域的限定纤维排列的3D COL1矩阵(厚度为130-250μm)的方案。该平台通过提供对用于细胞培养的微工程基质的直接访问，提供先进的细胞培养能力来对结构化组织微环境进行建模。
Aligned collagen I (COL1) fibers guide tumor cell motility, influence endothelial cell morphology, control stem cell differentiation, and are a hallmark of cardiac and musculoskeletal tissues. To study cell response to aligned microenvironments in vitro, several protocols have been developed to generate COL1 matrices with defined fiber alignment, including magnetic, mechanical, cell-based, and microfluidic methods. Of these, microfluidic approaches offer advanced capabilities such as accurate control over fluid flows and the cellular microenvironment. However, the microfluidic approaches to generate aligned COL1 matrices for advanced in vitro culture platforms have been limited to thin "mats" (<40 μm in thickness) of COL1 fibers that extend over distances less than 500 μm and are not conducive to 3D cell culture applications. Here, we present a protocol to fabricate 3D COL1 matrices (130-250 μm in thickness) with millimeter-scale regions of defined fiber alignment in a microfluidic device. This platform provides advanced cell culture capabilities to model structured tissue microenvironments by providing direct access to the micro-engineered matrix for cell culture.
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