Microchip-based 3D-Cell Culture Using Polymer Nanofibers Generated by Solution Blow Spinning.
Microchip-based 3D-Cell Culture Using Polymer Nanofibers Generated by Solution Blow Spinning.
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DOI：
10.1039/c7ay00756f
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发表时间：
2017-06-14
期刊：
影响因子：
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通讯作者：
Martin RS
中科院分区：
文献类型：
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作者：
Chen C;Townsend AD;Sell SA;Martin RS
聚合物纳米/微米纤维已经发现了许多应用，包括3D细胞培养和伤口敷料的产生。纤维可以通过包括静电纺丝的各种技术生产，其主要缺点包括需要高压电源（这可能导致诸如聚合物变性的问题）和缺乏便携性。最近，溶液吹塑纺丝（其中使用高速鞘气代替高电压）已被用于产生聚合物纤维。在这项工作中，我们使用吹塑纺丝来创建用于基于微芯片的3D细胞培养的纳米/微米纤维。首先，我们彻底研究了使用两种适合细胞培养的聚合物（聚己内酯，PCL和聚苯乙烯，PS）以及可能影响PCL和PS纤维质量的参数的3D打印气鞘装置的纤维生成。使用3D打印的鞘装置，发现鞘N2的压力和聚合物溶液的浓度决定了是否可以生产纤维以及所得的纤维形态。此外，我们展示了这些纤维如何通过直接将PCL纤维沉积在培养皿和孔板中来用于3D细胞培养。结果表明，PCL纤维与RAW264.7巨噬细胞具有良好的相容性，PCL纤维支架的厚度可达178 ± 14 μm。然后，将溶液吹塑纺丝（具有3D打印鞘装置）产生的PCL纤维与微流体装置首次整合，以制造具有流动组件的3D细胞培养支架。在流体装置上培养和刺激巨噬细胞后，发现整合的3D纤维支架是细胞外基质的更好模拟物（与平坦的2D基底相反），具有来自用脂多糖刺激的巨噬细胞的增强的亚硝酸盐积累（一氧化氮释放的产物）。还制备了PS纤维并将其整合到微流体装置中用于内皮细胞的3D培养，其保持存活至少72小时（在流动条件下为48小时）。这种方法将是有用的，涉及更现实的微芯片为基础的培养模型，研究细胞间的通信未来的研究。
Polymer nano/micro fibers have found many applications including 3D cell culture and the creation of wound dressings. The fibers can be produced by a variety of techniques that include electrospinning, the primary disadvantage of which include the requirement for a high voltage supply (which may cause issues such as polymer denaturation) and lack of portability. More recently, solution blow spinning, where a high velocity sheath gas is used instead of high voltage, has been used to generate polymer fibers. In this work, we used blow spinning to create nano/microfibers for microchip-based 3D cell culture. First, we thoroughly investigated fiber generation from a 3D printed gas sheath device using two polymers that are amenable to cell culture (polycaprolactone, PCL and polystyrene, PS) as well as the parameters that can affect PCL and PS fiber quality. Using the 3D printed sheath device, it was found that the pressure of the sheath N2 and the concentration of polymer solutions determine if fibers can be produced as well as the resulting fiber morphology. In addition, we showed how these fibers can be used for 3D cell culture by directly depositing PCL fibers in petri dishes and well plates. It is shown the fibers have good compatibility with RAW 264.7 macrophages and the PCL fiber scaffold can be as thick as 178 ± 14 μm. PCL fibers created from solution blow spinning (with the 3D printed sheath device) were then integrated with a microfluidic device for the first time to fabricate a 3D cell culture scaffold with a flow component. After culturing and stimulating macrophages on the fluidic device, it was found that the integrated 3D fibrous scaffold is a better mimic of the extracellular matrix (as opposed to a flat, 2D substrate), with enhanced nitrite accumulation (product of nitric oxide release) from macrophages stimulated with lipopolysaccharide. PS fibers were also made and integrated in a microfluidic device for 3D culture of endothelial cells, which stayed viable for at least 72 hours (48 hours under the flowing conditions). This approach will be useful for future studies involving more realistic microchip-based culture models for studying cell-to-cell communication.