Microfluidic Generation of Monodisperse, Structurally Homogeneous Alginate Microgels for Cell Encapsulation and 3D Cell Culture.
Microfluidic Generation of Monodisperse, Structurally Homogeneous Alginate Microgels for Cell Encapsulation and 3D Cell Culture.
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DOI：
10.1002/adhm.201500021
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发表时间：
2015-08-05
影响因子：
10
通讯作者：
Weitz DA
中科院分区：
文献类型：
--
作者：
Utech S;Prodanovic R;Mao AS;Ostafe R;Mooney DJ;Weitz DA
最近的研究表明，基本的细胞行为在二维和三维培养系统之间有很大的不同。为了确定这些根本差异的根源，设计可靠和精确控制的环境是必不可少的。虽然二维细胞培养是一种成熟的技术，但定义三维培养模型的制造仍然具有挑战性。我们提出了一种微流体生成微米尺寸三维细胞培养系统的新方法。我们使用触发离子交联形成生成高度单分散和结构均匀的海藻酸盐微珠。含有海藻酸盐和水溶性钙- edta复合物的混合物的水滴是由基于液滴的微流体形成的。在它们的络合形式下，钙离子均匀地分布在液滴内部，但无法进行交联过程。加酸是用来触发络合物的降解，释放钙离子的需求，可以物理交联藻酸盐链。这样就产生了一种均匀的水凝胶网络，它可以转移到水环境中而不失去其结构完整性。单个细胞可以被封装在这些受控的微环境中，提供结构支持，同时允许持续的营养供应。我们将单个间充质干细胞（MSCs）包封到微珠中，显示吸吸细胞的生长和增殖。
Recent studies have shown that basic cellular behavior varies significantly between two- and three-dimensional culture systems. To identify the origins of these fundamental differences the design of reliable and precisely controlled environments is essential. While 2D cell culture is a well-established technique, the fabrication of defined three-dimensional culture models is still challenging. We present a new method for the microfluidic generation of a micron-sized three-dimensional cell culture system. We use a triggered ionic crosslink formation to generate highly monodisperse and structurally homogeneous alginate microbeads. Aqueous droplets containing a mixture of alginate and a water-soluble calcium-EDTA complex are formed by droplet-based microfluidics. In their complexed form, the calcium ions are homogenously distributed inside the droplet but not accessible for the crosslinking process. Acid addition is used to trigger the degradation of the complex, releasing calcium ions on demand that can physically crosslink the alginate chains. A homogeneous hydrogel network is thus generated which can be transferred into an aqueous environment without losing its structural integrity. Single cells can be encapsulated into these controlled microenvironments which provide structural support while allowing for continuous nutrient supply. We encapsulate individual mesenchymal stem cells (MSCs) into the microbeads which show the aspired cell growth and proliferation.