| 题名 | 基于光诱导数字掩模的细胞操作与多维组装方法的研究 |
| 作者 | 杨文广 |
| 学位类别 | 博士 |
| 答辩日期 | 2017-11-29 |
| 授予单位 | 中国科学院沈阳自动化研究所 |
| 授予地点 | 沈阳 |
| 导师 | 王越超 ; 刘连庆 |
| 关键词 | 细胞操作与组装，水凝胶微制造，细胞行为学，组织工程，细胞多维信息获取 |
| 其他题名 | An optical induced method for cell manipulation and multidimensional assembly |
| 学位专业 | 模式识别与智能系统 |
| 中文摘要 | 本文开展基于光诱导数字掩模的细胞操作与多维组装方法的研究，旨在构建基于细胞多维信息的分析和研究体系，为多维细胞操作与组装的体外实现提供一种新的方法和解决思路。为了达到这个目标，本文开展了以下几个方面的研究工作：1）基于数字微镜阵列的紫外光诱导水凝胶制造方法。用于胞外环境构建的水凝胶制造系统是细胞多维信息获取与分析的关键，作者首先对紫外光引发自由基聚合反应进行相应的建模与仿真分析，从理论上分析了各种物质浓度及因素对于反应过程以及水凝胶制造结果的影响，阐明了光引发水凝胶微结构的制造机制。其次，构建了基于数字微镜阵列的紫外光诱导水凝胶加工系统，对搭建系统的光路进行了相应的仿真分析和优化处理，实现水凝胶个性化定制加工。使用本套系统加工水凝胶微结构可以在短短几秒钟内实现，展现出了效率高、灵活度高和重复性好的特点。最后，对水凝胶微结构的制造特性进行了研究，从实验上验证了各个参数对于所制造水凝胶结构机械特性的影响。2）细胞一维、二维图形化及细胞行为学的研究。在基于数字微镜阵列的紫外光诱导水凝胶制造系统的基础之上，作者通过制作水凝胶微结构来构建细胞的胞外环境，实现细胞的一维、二维图形化，并研究图形化对于细胞行为学的调控，其中包括以下几个方面：通过水凝胶微结构来调控细胞的形貌和机械特性，利用水凝胶微结构具有天然的生物惰性将细胞限制在特定环境中，来研究不同细胞的增殖特性。为了模拟癌细胞在人体血管内的迁移情况，利用水凝胶制造蜂窝状的沟道结构来研究不同细胞的迁移特性。为了解决细胞在PEGDA水凝胶表面不粘附的特性，作者通过在水凝胶中添加聚苯乙烯小球来改变水凝胶薄膜的表面粗糙度和机械特性，从而调控细胞对水凝胶薄膜的粘附特性。3）基于微坑阵列的三维细胞球状体模型建立及药物筛选。细胞是人体的基本组成单元，也是药物作用的主要对象，对于细胞的研究能够给细胞行为学和药物筛选带来极大的帮助。作者通过构建不同尺度和形状的微坑阵列，来实现对于三维球状体模型的构建，并进行了三维细胞球状体的分析和信息获取。此外，细胞球状体作为一种体外三维模型展现出了良好的耐药性，为了更加真实的模拟人体的环境，作者通过将光诱导水凝胶技术与微流控技术相结合，制作复合异质型细胞球状体，并进行了组合药物筛选。 4）微组织结构的高通量制造和三维模块化组装。面对药物筛选对人体微组织环境的需求，作者通过将微流控技术、光诱导水凝胶制造技术和光诱导介电泳技术进行有机的集成，提出了微小组织的在线制造和机器人同步装配策略（Organ Real-time Assembly on Chip），通过此方法能够根据需求在线制造不同种类的三维细胞微组织，并能同时采用微纳机器人技术进行在线组装，进而形成类人体生理环境的多细胞复杂组织连接体，为类人体生理环境的体外模拟提供了可行的解决方案。此外，整个过程采用机器人自动化方法实现，因而具备良好的可重复性和稳定性，从而保证了类人体生理构建的一致性，为未来组织再生和个性化药物筛选奠定了基础。 |
| 英文摘要 | The article focuses on developing an optical induced method for cell manipulation and multidimensional assembly based on digital micromirror device. In order to achieve the goal, this paper carry out the following research works: 1) Rapid fabrication of hydrogel microstructures using uv-induced DMD-based projection printing. Firstly, the free radical polymerization by UV irradiation is modeled and simulated. The influences of PEGDA concentration, TPO concentration, and UV laser exposure time on hydrogel fabricating results are analyzed theoretically. The manufacturing mechanism of hydrogel is elucidated. Secondly, an ultraviolet (UV)-curing method based on a digital micromirror device (DMD) for fabricating poly(ethylene glycol) diacrylate (PEGDA) hydrogel microstructures is presented. The simulation analysis and optimization of the optical path design of the building system are also carried out. By controlling UV projection in real-time using a DMD as digital dynamic mask instead of a physical mask, polymerization of the pre-polymer solution can be controlled to create custom-designed hydrogel microstructures. Arbitrary microstructures could also be fabricated within several seconds (<5 s) using a single-exposure, providing a much higher efficiency than existing methods, while also offering a high degree of flexibility and repeatability. Finally, the influences of parameters on mechanical properties are illustrated based on the study of characteristic of microstructure fabrication. 2) The study of 1D single cell and 2D cell patterns and cell behaviors regulated by the physical microenvironment constructed via hydrogel fabricating system. The hydrogel microstructures are used for constructing extracellular environment to achieve cell patterning. The shape effects on cellular growth and hybrid tissue layer were investigated by peeling off the microwells. And the surrounding microenvironment of breast cancer cells is constructed, enabling the investigation of the external environment’s effects on breast cancer cell behaviors. Breast cancer cell morphology and mechanical properties were modified by altering the microenvironment. Proliferation was higher in breast cancer as compared to normal cells, consistent with the primary characteristic of malignant tumors. Moreover, breast cancer cells migrated more rapidly when grown in a narrow channel as compared to a wider channel. To solve the inadhesion of cells to the surface of PEGDA film, the addition of PS into photo cross-linked PEGDA resulted in a hybrid film (PEGDA-PS) significantly alters surface morphology and mechanical properties and modulates the biocompatibility of the film. 3) 3D cell spheroids were established based on microwells. Cells are the basic elements of body and targets of drug treatment. Cell spheroids (3D) can be formed using microwell array and the analysis of biological characteristics are achieved separately. Furthermore, 3D multicellular spheroids were successfully established in a controllable and highthroughput format. Our method facilitates the formation of 3D spheroids, and the growth and drug response of these spheroids can be monitored in real time. Overall, the 50% inhibition concentration of 3D spheroids is more than twice that of cells cultured on a 2D monolayer, which highlights the importance of the 3D spheroids for more closely simulating the parent tissue condition in vivo. 4) High-throughput fabrication and modular assembly of 3D heterogeneous microscale tissues. Cell monolayer is different from human body, which results in a loss of information during drug screening. To solve this problem, high-throughput fabrication and modular assembly of 3D heterogeneous microscale tissues is presented. And a versatile method for fabricating customized microstructures and reorganizing building blocks composed of functional components into a combined single geometric shape is demonstrated. The arbitrary microstructures are dynamically synthesized in a microfluidic device and then transferred to an optically induced electrokinetics chip for manipulation and assembly. Moreover, building blocks containing different cells can be arranged into a desired geometry with specific shape and size, which can be used for microscale tissue engineering. As a whole, the fabrication and assembly methodology with repeatability and stability developed here holds great promise as a platform for creating artificial functional biological architectures for use in drug screening, regenerative medicine, and tissue transplantation. |
| 语种 | 中文 |
| 产权排序 | 1 |
| 页码 | 114页 |
| 内容类型 | 学位论文 |
| 源URL | [http://ir.sia.cn/handle/173321/21278]  |
| 专题 | 沈阳自动化研究所_机器人学研究室 |
| 推荐引用方式 GB/T 7714 | 杨文广. 基于光诱导数字掩模的细胞操作与多维组装方法的研究[D]. 沈阳. 中国科学院沈阳自动化研究所. 2017. |
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