静电纤维聚合物支架用于组织工程和细胞培养
Summary
静电聚合物组织工程和细胞培养的过程中，在这篇文章中解决。具体来说，静电photopatterning多聚合物静电额外的处理能力的光反应macromers描述。
Abstract
作为组织工程领域的的发展，是一个巨大的需求，生产更合适的材料和加工技术，以解决更复杂的器官和组织的要求（如机械和血管）。静电是一种流行的技术来创造，模仿原生细胞外基质的结构和大小规模的纤维支架。这些纤维支架也有用的，因为细胞培养基质的纤维可以被用来直接细胞的行为，包括干细胞的分化（见广泛的审查由莫克<em>等。</em>和西尔<em>等。</em>了解更多信息）。在这篇文章中，我们描述了静电纺丝聚合物的一般过程，作为一个例子，electrospin一个反应的光线照射交联透明质酸（见Ifkovits<em>等。</em>审查photocrosslinkable材料）。我们还介绍了进一步的处理能力，如photopatterning多聚合物支架形成。 Photopatterning可用于创建渠道和多尺度孔隙支架，以增加细胞浸润和组织分布。多聚合物支架是有用的，以便更好地调整棚架的属性（力学和退化），其中包括专为细胞浸润的孔隙度。此外，这些技术可以扩展到包括广泛的聚合物和反应macromers的创建提供必要的线索，开发成功的组织工程结构的复杂支架。
Protocol
答：单聚合物静电准备电的解决方案之前，在光引发剂0.5％的溶液，Irgacure 2959（I2959），去离子水溶解在37 ° C几天。这一步是没有必要的，如果光反应的聚合物不被使用。 结合methacrylated透明质酸（梅阿，看到伯迪克合成等 ），聚（环氧乙烷）（PEO，900 kDa的），在去离子水I2959准备与终浓度为2％，3％梅阿的解决方案私立教育机构，和0.05 wt％的I2959。使用一个旋涡混合解决方…
Discussion
静电是用来准备从聚合物纤维支架。 Photocrosslinkable支架的基础上被用来作为一个例证，在光线照射是需要交联透明质酸。随着反应macromers的使用，如梅阿，先前已证明增强细胞分布的通道被纳入棚架光交联过程中使用口罩，以形成宏观和微观多孔支架。此外，两个不同的聚合物，同时静电使用我们自定义的仪器。两种截然不同的纤维人口的存在，验证与荧光染料除了每个静电解决方案，后来用荧…
Acknowledgements
这项工作是由美国心脏协会Predoctoral的奖学金JLI和国家机构的希思授予R01AR056624支持。
Materials
| Material Name | Type | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| DAPI | Reagent | Invitrogen | D1306 | |
| I2959 | Reagent | Ciba Specialty Chemicals | ||
| PEO 200 kDa | Polysciences | 17503 | ||
| PEO 900 kDa | Reagent | Sigma | 189456 | |
| Methacryloxethyl thiocarbamoyl rhodamine B | Reagent | Polysciences | 23591-100 | Prepare stock solution in DMSO |
| Live/Dead Stain Kit | Reagent | Invitrogen | L3224 | Contains Calcein (stains live cells green) and ethidium homodime (stains red dead cells) |
| Syringe Pump | Equipment | KD Scientific | KDS100 | Two are needed for dual polymer spinning |
| Power Source | Equipment | Gamma High Voltage | ES30P-5W | Two are needed for dual polymer spinning |
| Motor | Equipment | Triem Electric Motors, Inc | 0132022-15 | Must attach to a custom built mandrel |
| Tachometer | Equipment | Network Tool Warehouse | ESI-330 | Use to monitor mandrel speed |
| Omnicure UV Spot Cure System with collimating adapter | Equipment | Exfo Life Sciences Division | S1000 | |
| Silicone Tubing | Equipment | McMaster-Carr | 51135K151 | |
| Luer Lock Female Adapter | Equipment | McMaster-Carr | 51525K293 | |
| Luer Lock Male Adapter | Equipment | McMaster-Carr | 51525K143 | |
| Needles | Equipment | Fisher Scientific | 14-825-16H | |
| Coverslips | Equipment | Corning | 2875-22 |
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Tags
ElectrospinningFibrous Polymer ScaffoldsCell CultureElectrospinning TechniqueNative Extracellular MatrixCell Culture SubstratesStem Cell DifferentiationHyaluronic AcidCrosslinkingLight ExposurePhotocrosslinkable MaterialsPhotopatterningMulti-polymer Scaffold FormationChannelsPorosityCellular InfiltrationPolymersReactive Macromers
Cite This Article
Ifkovits, J. L., Sundararaghavan, H. G., Burdick, J. A. Electrospinning Fibrous Polymer Scaffolds for Tissue Engineering and Cell Culture. J. Vis. Exp. (32), e1589, doi:10.3791/1589 (2009).