细胞高通量捕获和排列方法的最新进展

Latest advances in high-throughput cell capture and arrangement

背景:细胞是生命体活动的基本单位,理解生命体过程中的规律,需要以研究细胞为基础来深入探索细胞间的联系和行为。大多数的生物测定都是基于大细胞群体,但是其缺点是无法忽略细胞异质性的影响,并且失去了在细胞应答过程中重要的瞬时数据。目前对实现单细胞捕获和诱导体外排列来研究细胞具体行为的方法缺乏系统性的表述和研究。目的:总结并讨论微流控技术、微接触印刷技术及利用力学、电学及磁学来制作细胞捕获陷阱的方法及实现后续排列、生长、铺展、分化控制的可行性和最新进展。方法:作者检索1995至2017年百链云数据库和Pub Med数据库,纳入与单细胞捕获及筛选微加工方法相关的文献,并进行系统整理、归纳总结和分析。结果与结论:共检索到文献241篇,按照纳入和排除标准筛选后,共纳入35篇文献。通过阅读、总结和分析,结果表明:随着微纳米级加工技术的逐步成熟及它在细胞生物学和医学研究中的逐步深入,微流控技术和微接触印刷技术既是目前研究的重点又被很多研究者进行了一定程度的改进,除此之外一些新的材料也逐步应用。微流控技术在细胞捕获率方面有着领先的优势,而改进后的微接触印刷技术更注重细胞的后续铺展及排列。在不损害细胞活性的前提下,精准的细胞捕获和排列以及合适的后续培养环境,对引导细胞的铺展、融合及分化方面至关重要。

BACKGROUND： Cells are the basic units of all life activities. In order to grasp the law of life process, it is essential to explore intercellular interactions and cell behaviors. Most current biological assays in large cell populations ignore the effects of cell heterogeneity and lose important temporal data in the process of averaging cellular responses. High-throughput single-cell capture and arrangement are of great significance to the research on cell biology. However, to date, there has no systematic study and description of the methods for cell capturing and alignment in vitro. OBJECTIVE： To summarize the methods of microfluidics technology, surface topographical technology and various traps based on mechanics, magnetics, and electrophoretic sorting to spatially collect single cells so as to discuss the feasibility and the latest progress of cell shape control and cell alignment. METHODS： The authors performed a data retrieval of Pub Med and Bailianyun databases to search the articles（1995-2017） addressing the single cell capture and alignment in vitro and reviewed the literatures systematically. RESULTS AND CONCLUSION： A total 241 articles were retrieved, and 35 articles were finally involved in the analysis according to the inclusion and exclusion criteria. After summarizing and analyzing, the results indicated that microfluidics, micro-contact printing, micro-well arrays, micro-pore membrane, electrical stimulation and magnetic deflection are commonly used in cell capture and functional assay. With the development of micro-scale technologies and in-depth research of cell behavior, microfluidic technology and micro-contact printing technology have become a hot topic of research with certain improvement in the capture efficiency. In addition, some new materials are gradually developed and applied. Microfluidic technology has a leading advantage in cell capture rate, while the improved micro-contact printing technology mostly concerns subsequent cell spreading and alignment. Under the proper culture environment, precise cell capture and alignment are essential to guide cell spreading, fusion, and differentiation.