Name: construction of modular hydrogel sheets for micropatterned macro-scaled 3d cellular architecture
Uploaded: 2016-01-11T10:30:00
Description: Watch this Scientific Journal Video about Construction of Modular Hydrogel Sheets for Micropatterned Macro-scaled 3D Cellular Architecture at JoVE.com

This research was supported by a National Leading Research Laboratory Program (Grant NRF-2013R1A2A1A05006378) through the National Research Foundation of Korea funded by the Ministry of Science, ICT and Future Planning. The authors also acknowledge a KAIST Systems Healthcare Program.

1.制备微模式化模具和水凝胶的<ol><li>通过标准的两步光刻技术15,17用于铸造PDMS模具生产使用的SU-8光致抗蚀剂的硅晶片的表面上所希望的微观图案。示出的例子使用一个肝小叶状网状图案（ 图1）。 </li><li>称出PDMS和固化剂溶液与1：5的比例（即，12.5克PDMS和2.5克固化剂）。 </li><li>彻底混合15克的溶液，脱气气泡在真空室中，然后蔓延混合溶液到硅晶片的微图案化表面?...

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这个协议提供了一种制造模块化的水凝胶片，并把它们组装，以形成三维的细胞支架的一个简单方法。 为了构建在短时间内鲜明图案化藻酸盐的结构，我们应确定一个交联的过程，可以创造足够刚性的结构，以从模具保持复杂微图案，以及维持细胞活力和新陈代谢。我们已经开发出一种交联方法，包括溶胶 - 凝胶转变，喷使用以雾状形式的加湿器的交联试剂。如果没有这种?...

水凝胶是特别有前途的生物材料，并且预计将在基础生物学，药理学试验和医学重要1的水凝胶为基础的蜂窝构造的Biofabrication已经建议减少使用的动物实验中，2,3-取代移植组织，4和提高基于细胞的测定。5,6-含有水（加氢）的粘弹性材料（凝胶）允许进行封装和保持在支架结构，以控制3D细胞微环境的大量细胞。与微流体或微图案的技术的指导组合，该水凝胶结构的几何形状可以精确地在细胞比例控制。迄今为止，多种水凝胶的形状，包括颗粒，7 - 9纤维，10 - 12，片材，13 - 15已经被用作建筑物单元在自下而上的Appro酸痛到宏观规模的多细胞结构的制造。 既基于水凝胶的颗粒和纤维一直容易且迅速地制造为应用程序作为微尺度蜂窝环境中，使用微流体装置的流体控制。然而，如工程化组织的基本单位，它将是复杂的重新排列，并扩大其体积为宏观结构。16，是更难以实现宏观规模构建体相比，以产生微米级的基本模块。可用于水凝胶基的构建体的片状单元通过简单的组装过程来增加支架的体积。必然，水凝胶片材的堆叠层不仅提供一个容积增加，而且在一个三维空间中的几何延伸。 我们以前曾报道制造微图案化的水凝胶片材，13的方法- 15连同其组装成多捻ERED蜂窝架构。该技术使得复杂微图案和蜂窝结构的模块化设计通过多层结构的层叠过程。通过堆叠模块化水凝胶片，这是微图案的制造中，可以实现一个三维细胞培养系统具有受控的宏观尺度细胞微环境。此视频协议描述了一个简单而强大的制造方法，可用于构建模块化水凝胶片材，基于人肝癌细胞系（HepG2细胞）。我们证明本文简单操作这些图案化的模块化的水凝胶片材，和它们的组装成的多层结构。

<table border="0" cellpadding="0" cellspacing="0" width="958">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:288px;">Sylgard 184 Silicone Elastomer Kit</td>
			<td style="width:111px;">Dow Corning Corporation</td>
			<td style="width:163px;">000000000001064291</td>
			<td style="width:397px;"></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:288px;">Pluronic F-127</td>
			<td style="width:111px;">Sigma-Aldrich</td>
			<td style="width:163px;">P2443</td>
			<td>Powdered nonionic surfactant&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Alginic acid sodium salt, low viscosity</td>
			<td style="width:111px;">Alfa Aesar</td>
			<td style="width:163px;">B25266</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Calcium chloride dihydrate</td>
			<td style="width:111px;">Sigma-Aldrich</td>
			<td style="width:163px;">C7902</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Ultrasonic humidifier</td>
			<td style="width:111px;">MediHeim</td>
			<td style="width:163px;">MH-2800</td>
			<td>Modified equipment, Maximum sprayed rate: 250 mL/h</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:288px;">Nylon net filter hydrofilic, 180 &#956;m</td>
			<td style="width:111px;">EMD Millipore</td>
			<td style="width:163px;">NY8H04700</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:288px;">Polycarbonate mold</td>
			<td style="width:111px;"></td>
			<td style="width:163px;"></td>
			<td>Customized mold for fabrication of a PDMS frame pattern</td>
		</tr>
	</tbody>
</table>

construction of modular hydrogel sheets for micropatterned macro-scaled 3d cellular architecture

水凝胶可以在使用微流体或微图案的技术，以提供在体内样的三维（3D）几何组织微观尺度进行图案化。所得三维水凝胶为基础的蜂窝结构已被引入作为替代动物实验为先进的生物研究，药理学试验和器官移植的应用程序。尽管水凝胶为基础的颗粒和纤维，可以容易地制造，这是很难操纵它们用于组织重建。在这种视频中，我们描述了一种制造方法，微图案化藻酸盐水凝胶片材，以及它们的组件，以形成一个宏观尺度三维细胞培养体系具有受控细胞微环境。 200微米，具有精确的微图案 - 采用钙胶凝剂薄雾形式，薄水凝胶片材易于与厚度在100〜生成。细胞然后可与水凝胶片材在几何指导培养独立的条件。此外，水凝胶片材可以容易地使用微量具有端切割尖端操纵，并且可以通过使用图案化的聚二甲基硅氧烷（PDMS）帧堆叠它们组装成多层结构。这些模块化水凝胶片，其可以使用一个浅显工艺制造，具有的体外药物分析和生物学研究，包括微观和宏观结构和组织重建的功能研究潜在的应用。

我们已经描述了制造和操纵独立的细胞水凝胶表。 如图1中，我们制备微图案化的PDMS模具，和包含细胞的水凝胶上样到这些模具的亲水性表面和交联使用增湿以产生胶凝剂的气溶胶雾。以下从模具释放，HepG2细胞都在独立的凝胶片材具有各种图案（图2）中培养。因此，该薄水凝胶片提供了一种三维培养环境。此外，模块化的水凝胶片材可以通过层...

Watch this Scientific Journal Video about Construction of Modular Hydrogel Sheets for Micropatterned Macro-scaled 3D Cellular Architecture at JoVE.com

Construction of Modular Hydrogel Sheets for Micropatterned Macro-scaled 3D Cellular Architecture

We describe the fabrication of micropatterned hydrogel sheets using a simple process, which can be assembled and manipulated in a freestanding form. Using these modular hydrogel sheets, a simple macro-scaled 3D cell culture system can be generated with a controlled cellular microenvironment.

构建模块化水凝胶片的微模式化宏观缩放3D细胞结构

Hydrogels can be patterned at the micro-scale using microfluidic or micropatterning technologies to provide an in vivo-like three-dimensional (3D) tissue ...

Research

JoVE Journal

Bioengineering

Tissue Engineering: Construction of a Multicellular 3D Scaffold for the Delivery of Layered Cell Sheets

组织工程：建设多细胞三维支架的层状细胞片交货

Many tissues, such as the adult human hearts, are unable to adequately regenerate after damage.2,3 Strategies in tissue engineering propose innovations to ...

科研

生物工程

The Encapsulation of Cell-free Transcription and Translation Machinery in Vesicles for the Construction of Cellular Mimics

无细胞的转录和翻译的机械囊泡的封装建设模拟蜂窝

As interest shifts from individual molecules to systems of molecules, an increasing number of laboratories have sought to build from the bottom up ...

3D Hydrogel Scaffolds for Articular Chondrocyte Culture and Cartilage Generation

3D水凝胶支架的关节软骨细胞培养及软骨生成

Human articular cartilage is highly susceptible to damage and has limited self-repair and regeneration potential. Cell-based strategies to engineer ...

Gradient Strain Chip for Stimulating Cellular Behaviors in Cell-laden Hydrogel

在单元格拉丹水凝胶中刺激细胞行为的应变梯度芯片

Artificial guidance for cellular alignment is a hot topic in the field of tissue engineering. Most of the previous research has investigated single ...

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

改良的3D 水凝胶培养的原发胶质细胞的体外Neuroinflammation 模型的建立

In the central nervous system, numerous acute injuries and neurodegenerative disorders, as well as implanted devices or biomaterials engineered to enhance ...

Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System

3D 动态培养系统构建多层间充质干细胞片材

Stem cell therapy shows a promising future in regenerating injured organ and tissues, and the cell sheet technique has been developed to improve the low ...

3D Analysis of Multi-cellular Responses to Chemoattractant Gradients

趋化梯度多细胞响应的三维分析

Various limitations of 2D cell culture systems have sparked interest in 3D cell culture and analysis platforms, which would better mimic the spatial and ...

3D Printed Porous Cellulose Nanocomposite Hydrogel Scaffolds

三维印刷多孔纤维素纳米复合水凝胶脚手架

This work demonstrates the use of three-dimensional (3D) printing to produce porous cubic scaffolds using cellulose nanocomposite hydrogel ink, with ...

Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks

明胶美沙克里洛醇水凝胶生物油墨3D生物印刷方案

Gelatin methacryloyl (GelMA) has become a popular biomaterial in the field of bioprinting. The derivation of this material is gelatin, which is hydrolyzed ...

Simple, Affordable, and Modular Patterning of Cells using DNA

使用DNA对细胞进行简单、经济、模块化的图案化

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells ...