Creation of highly functional thermal and fluids mechanism for micro cell processing

本研究的目的是开发一种新型的微细胞处理系统，利用微尺度中固有的热和流体现象，在微通道中高效地混合、标记、分类和传输细胞。作为一种特定的应用，假定使用成体干细胞的再生医学，并已开发出能够有效地从细胞混合物中提取稀有细胞的细胞分选系统。更具体地说，为了理解微通道中细胞/粒子的运动，已经发展了高精度的数值格式和新的成像方法。通过采用新型生物兼容材料的MEMS制造技术的发展和对原型微细胞分选器的实验评估，得出了可用于未来临床应用的细胞分选系统的具体设计指导原则。我们发现，与以前的微型细胞分选器相比，该方法的吞吐量更高，并且采用向上计数的方法可以处理相对大量的临床所需的细胞样本。此外，还提出了一种新的基于黏附的细胞分选系统，该系统利用抗原-抗体在壁上的相互作用来减慢细胞滚动的速度，并开发了其原型。使用该系统，可以消除对目标细胞的标记。我们还提出了一种新的氨基功能化的抗体固定化方法，传统的细胞分选/孵育过程耗时长，需要大量的人力资源。本研究提出的基于微尺度热流特性的细胞分选方法，将降低细胞分选的成本和分析时间，最终为再生医学的发展做出贡献。

The objective of the present study is to develop a novel micro cell processing system for efficient mixing, labeling, sorting and transporting of cells in micro channels by using heat and fluid phenomena inherent in microscale. As a specific application, regenerative medicine using adult somatic stem cells is assumed, and cell sorting systems that can efficiently extract rare cells from cell mixture have been developed. More specifically, high-precision numerical scheme and a novel imaging methods have been developed for understanding cell/particle motions in micro channels. Specific design guiding principle for cell sorting system that can be used for future clinical application is obtained by development of MEMS fabrication technologies using new biocompatible materials and experimental evaluation of the prototype micro cell sorters.A lamination micro mixer and an immunomagnetic cell sorting system using the mixers have been developed. We found that the throughput can be higher than previous micro cell sorters, and that processing relatively-large amount of cell samples needed for clinical use is possible when the numbering-up approach is adopted. In addition, novel adhesion-based cell sorting systems that utilize deceleration of cell rolling by antigen-antibody interaction on the wall has been proposed, and its prototype is developed. With this system, one can eliminate labeling of the target cells. We also propose a new immobilization method of antibody with amino-functionalized parylene.Conventional cell sorting/incubation process is time consuming and requires a long time and much human resources. Cell sorting methods proposed in the present study based on characteristics of microscale heat and fluid flows will reduce cost and analysis time of cell sorting, and eventually make contribution to advances of regenerativie medicine.