微流控芯片系统在循环肿瘤细胞分离检测中的应用进展

循环肿瘤细胞(CTCs)的分离分析一直是肿瘤相关研究中的热点方向,作为液体活检的重要标志物之一,其在外周血中的含量与癌症病发状况密切相关.然而人体血液中CTCs的含量非常低,通常来说仅有0～10个/mL,因此在开展临床血液样本中CTCs的检测前,往往需要对样本进行前处理,以实现CTCs的分离和富集.微流控芯片技术凭借样...     

微流控芯片分选富集循环肿瘤细胞的研究进展

近年来,循环肿瘤细胞(CTCs)研究得到了越来越多的关注,许多研究报告已经证实其在肿瘤转移的早期诊断、治疗方案选择、个体化治疗及探索肿瘤转移机制等方面具有潜在的价值,然而CTCs在循环系统中的含量极低,这成为限制其临床相关应用的主要难点。微流控芯片技术具有低成本、快速、高通量及操作简单等优势,利用微流控芯片可实现CTCs的高速、高回收率、高纯度的分选富集,近年来得到广泛的关注。本文综述了近年来在微流控芯片内进行CTCs分选富集的研究并探讨了各种方法的优缺点,并在本研究团队的研究基础上进行了展望。     

微流控芯片分选临床样品中循环肿瘤细胞的研究进展

癌症作为常见病正严重威胁着我国乃至全球居民的健康。循环肿瘤细胞(CTCs)是一类由癌变部位释放并进入血液中的癌细胞,其在癌症的早期诊断、个体化及肿瘤转移机制研究等方面的作用正逐渐被发现和认可,但由于血液中的CTCs含量极少,对其分选极具挑战。微流控芯片作为一种微型化、高通量、集成化平台,在CTCs研究中彰显了独特的优势,相关报道也越来越多。随着研究的深入,微流控芯片技术不再局限于基于模型样品的方法学开发,而是更注重于能否用于临床实际样品中CTCs的检测,但目前未见该角度的综述报道。为此,文章综述了近年来用于临床实际样品CTCs分析的微流控芯片分选技术,并探讨了微流控芯片用于CTCs分选的发展趋势。     

微流控技术在循环肿瘤细胞异质性分析领域的应用

循环肿瘤细胞(circulating tumor cells,CTCs)的检测是液体活检的重要组成部分,可以通过低损伤甚至无创的方法达到检测体液中肿瘤细胞的目的 .肿瘤细胞在增殖、转移过程中极易产生多种类型的变异,如基因突变、蛋白质组学和表观遗传学改变等,这导致肿瘤细胞个体间发生差异,给临床诊疗带来巨大挑战.通过CTC...     

基于微流控技术的循环肿瘤细胞分选研究

循环肿瘤细胞(CTCs)出现于癌症患者的外周血中,是一种重要的游离态组织样本,对于癌症的早期诊断和预后评估具有非常重要的临床诊断价值。由于血液中CTCs含量极少,对其进行分选富集是CTCs检测和分析的一个重要预处理步骤。传统的宏观方法虽然也能实现细胞分离,但存在耗时长、样品需求量大、目标细胞损失严重及硬件设备依赖性高等不足。近年来兴起的微流控技术可在微米尺度范围内集成物理、化学及生物手段,易于实现整体器件的微型化和低成本便携式发展,为稀有CTCs的高灵敏度、高效分选提供重要的潜在技术手段。本文综述了微流控技术实现CTCs分选的最新研究进展,详细阐述了各种被动、主动分选方法的原理及成功应用实例,分析各方法的优缺点,提出一种新型的多级分选芯片结构,并最后探讨了微流控CTCs分选芯片在临床应用中面临的挑战及未来的发展趋势。     

基于流体动力单细胞高效捕获的微流控芯片结构研究进展

单细胞分析对于重大疾病的早期诊断及治疗、药物筛选和生理病理过程的研究具有重要意义。微流控芯片能够精确控制单细胞的微环境,实时监测单细胞的行为,已成为单细胞分析的强大工具。单细胞捕获是单细胞分析的重要步骤。目前已报道了多种微流控芯片用于单细胞捕获的方法,其中基于流体动力的微流控芯片单细胞捕获方法具有操作方便、单细胞捕获效率高等优点,受到研究人员的广泛关注及使用。为了全面了解基于流体动力的微流控芯片单细胞捕获方法的研究现状,掌握单细胞高效捕获的微流控芯片结构设计,实现单细胞精准快速分析,本文综述了基于流体动力的单细胞高效捕获（>70%）原理及微流控芯片结构,根据结构设计不同分为微井结构、微柱结构和旁路通道结构,介绍了单细胞高效捕获的微流控芯片优化过程,总结了微流控芯片的材质、结构特点及单细胞捕获效率等,对不同单细胞捕获结构的优势及不足进行了分析。最后,对基于流体动力的微流控芯片单细胞捕获方法的发展趋势进行了展望。     

微流控芯片细胞实验室

以作者所在课题组近年开展的研究工作为基础,阐述了微流控芯片细胞实验室的平台特征,并从细胞个体、群体和多细胞生命体研究等三个方面概述微流控芯片细胞实验室的应用对象特征,显示其在生物医学领域的应用前景。     

微流控芯片的研究及产业化

以大连研究团队的近期工作为基础,结合2015年末召开的“深圳-大连微流控芯片及其产业化战略研讨会”内容,扼要阐述作者对近期微流控芯片的研究及产业化的基本看法.鉴于微流控芯片研究的主流已从平台构建和方法发展转为不同领域的广泛应用,本文重点介绍了微流控芯片在现代生物化学分析、即时诊断、材料筛选-材料合成以及组织-器官仿生等4个应用领域的研究趋势,讨论了3D打印技术的崛起对微流控芯片的影响和挑战,阐述了微流控芯片作为当代极为重要的新兴科学技术平台和国家层面产业转型的潜在战略领域,在全球范围内产业化的发展势头.全文引用文献69篇.     

微流控芯片二维电泳研究进展

综述了微流控芯片二维电泳技术及其在生命科学中的应用,包括胶束电动力学毛细管色谱(MEKC)与毛细管区带电泳(CZE)、等电聚焦(IEF)与CZE、开管电色谱(OCEC)与CZE耦联等模式的二维微流控芯片。展望了二维微流控芯片的应用前景。     

微流控芯片子宫

由于体外环境对受精和胚胎发育的影响,现有人工辅助生殖技术存在受精成功率低和胎儿出生后风险高的问题。针对上述问题,本研究发展了一种基于微流控芯片的人工子宫,其特色在于使用子宫内膜细胞-胚胎共培养机制,并以连续灌流方式实现细胞与外界物质交换。实验利用上述芯片完成了排卵、受精、着床以及胚胎发生等一系列过程。与传统方法相比,芯片方法不仅操作简便,而且可以获得更高的桑椹胚率和囊胚率。已经取得的研究结果显示,这种芯片子宫有希望发展成人工辅助生殖的有力工具。     

DNA Nanolithography Enables a Highly Ordered Recognition Interface in a Microfluidic Chip for the Efficient Capture and Release of Circulating Tumor Cells

Microfluidic chips with nano‐scale structures have shown great potential, but the fabrication and cost issues restrict their application. Herein, we propose a conceptually new “DNA nanolithography in a microfluidic chip” by using sub‐10 nm three‐dimensional DNA structures (TDNs) as frameworks with a pendant aptamer at the top vertex (ApTDN‐Chip). The nano‐scale framework ensures that the aptamer is in a highly ordered upright orientation, avoiding the undesired orientation or crowding effects caused by conventional microfluidic interface fabrication processes. Compared with a monovalent aptamer modified chip, the capture efficiency of ApTDN‐Chip was enhanced nearly 60 % due to the highly precise dimension and rigid framework of TDNs. In addition, the scaffolds make DNase I more accessible to the aptamer with up to 83 % release efficiency and 91 % cell viability, which is fully compatible with downstream molecular analysis. Overall, this strategy provides a novel perspective on engineering nano‐scaffolds to achieve a more ordered nano‐topography of microfluidic chips.     

Modification of Hemodialysis Membranes for Efficient Circulating Tumor Cell Capture for Cancer Therapy

Background: It is well known that more than 90% of cancer deaths are due to metastases. However, the entire tumorigenesis process is not fully understood, and it is evident that cells spreading from the primary tumor play a key role in initiating the metastatic process. Tumor proliferation and invasion also elevate the concentration of regular and irregular metabolites in the serum, which may alter the normal function of the entire human homeostasis and possibly causes cancer metabolism syndrome, also referred to as cachexia. Methods: We report on the modification of commercially available hemodialysis membranes to selectively capture circulating tumor cells from the blood stream by means of immobilized human anti-EpCAM antibodies on the inner surface of the fibers. All critical steps are described that required in situ addition of the immuno-affinity feature to hemodialyzer cartridges in order to capture EpCAM positive circulating tumor cells, which represents ~80% of cancer cell types. Results: The cell capture efficiency of the suggested technology was demonstrated by spiking HCT116 cancer cells both into buffer solution and whole blood and run through on the modified cartridge. Flow cytometry was used to quantitatively evaluate the cell clearance performance of the approach. Conclusions: The suggested modification has no significant effect on the porous structure of the hemodialysis membranes; it keeps its cytokine removal capability, addressing cachexia simultaneously with CTC removal.     

微流控芯片电泳快速分离脂蛋白

描述了一种芯片电泳快速分离脂蛋白的方法. 利用自制的微流控芯片及激光诱导荧光技术电泳分离经硝基苯并噁二唑-C₆-酰基鞘胺醇预染的脂蛋白标本, 在40 mmol/L tricine缓冲液(pH 9.4)中加入40 mmol/L甲基葡胺, 在500 V电压下40 s进样, 在2000 V 电压下2 min内完成分离, 可出现低密度脂蛋白(LDL)与高密度脂蛋白(HDL)两条脂蛋白区带, 5次重复性试验其出峰时间变异系数(CV)为2.6%. 本法为高血脂患者提供了一种快速、简便、灵敏、重复性好的诊断方法.     

Beyond the Capture of Circulating Tumor Cells: Next‐Generation Devices and Materials

Over the last decade, significant progress has been made towards the development of approaches that enable the capture of rare circulating tumor cells (CTCs) from the blood of cancer patients, a critical capability for noninvasive tumor profiling. These advances have leveraged new insights in materials chemistry and microfluidics and allowed the capture and enumeration of CTCs with unprecedented sensitivity. However, it has become increasingly clear that simply capturing and counting tumor cells launched into the bloodstream may not provide the information needed to advance our understanding of the biology of these rare cells, or to allow us to better exploit them in medicine. A variety of advances have now emerged demonstrating that more information can be extracted from CTCs with next‐generation devices and materials featuring tailored physical and chemical properties. In this Minireview, the last ten years of work in this area will be discussed, with an emphasis on the groundbreaking work of the last five years, during which the focus has moved beyond the simple capture of CTCs and gravitated towards approaches that enable in‐depth analysis.     

纳米生物无机界面的构筑与循环肿瘤细胞分离

纳米生物无机界面的研究是无机化学学科新兴的前沿领域之一。纳米结构的无机材料在仿生界面、细胞界面、生物检测界面等领域扮演着越来越重要的角色。近几年来,无机纳米结构被尝试用于痕量循环肿瘤细胞(Circulating Tumor Cells,CTCs)分离的基础探索研究中,并展现出非常吸引人的应用前景。痕量CTCs的高效分离对于癌症早期检测、术后监测及生物学研究等具有重要的意义。本文主要综述纳米生物无机界面在CTCs分离中的应用,详细介绍其发展现状,并对未来做一展望。     

纳米生物无机界面的构筑与循环肿瘤细胞分离

纳米生物无机界面的研究是无机化学学科新兴的前沿领域之一。纳米结构的无机材料在仿生界面、细胞界面、生物检测界面等领域扮演着越来越重要的角色。近几年来, 无机纳米结构被尝试用于痕量循环肿瘤细胞(Circulating Tumor Cells, CTCs)分离的基础探索研究中, 并展现出非常吸引人的应用前景。痕量CTCs的高效分离对于癌症早期检测、术后监测及生物学研究等具有重要的意义。本文主要综述纳米生物无机界面在CTCs分离中的应用, 详细介绍其发展现状, 并对未来做一展望。     

Spectrally Combined Encoding for Profiling Heterogeneous Circulating Tumor Cells Using a Multifunctional Nanosphere-Mediated Microfluidic Platform

Comprehensive phenotypic profiling of heterogeneous circulating tumor cells (CTCs) at single-cell resolution has great importance for cancer management. Herein, a novel spectrally combined encoding (SCE) strategy was proposed for multiplex biomarker profiling of single CTCs using a multifunctional nanosphere-mediated microfluidic platform. Different cellular biomarkers uniquely labeled by multifunctional nanosphere barcodes, possessing identical magnetic tags and distinct optical signatures, enabled isolation of heterogeneous CTCs with over 91.6 % efficiency and in situ SCE of phenotypes. By further trapping individual CTCs in ordered microstructures on chip, composite single-cell spectral signatures were conveniently and efficiently obtained, allowing reliable spectral-readout for multiplex biomarker profiling. This SCE strategy exhibited great potential in multiplex profiling of heterogeneous CTC phenotypes, offering new avenues for cancer study and precise medicine.     

一种用于乳酸检测的新型微流控芯片

基于光度吸收原理,设计了一种集成片上混合和光纤检测功能于一体的微流控芯片,用于细胞和组织培养过程中乳酸代谢浓度的在线检测。通过光学设计软件(Zem ax)优化设计了吸收光路,得到微流控芯片沟道宽度为250μm,利用计算流体动力学软件(CFD)模拟确定了乳酸和显色剂片上混合时微流控芯片沟道的溶液完全混合位置和光纤检测点,采用微电子机械系统(MEMS)加工了基于聚二甲基硅氧烷(PDMS)的微流控芯片,将片上混合和光纤检测功能集成在一个以PDMS和载玻片组成的芯片上。实验结果表明,该芯片成功实现了乳酸和显色剂的片上混合和实时检测,检出限(LOD)为0.52 mmol/L(47 mg/L),乳酸浓度从1 mmol/L变化至5 mmol/L时的芯片响应时间为130 s,能够满足细胞和组织培养过程中乳酸在线检测要求。     

Construction of Tumor Tissue Array on An Open-Access Microfluidic Chip

An open-access microfluidic chip which enabled automatic cell distribution and complex multi-step operations was developed. The microfluidic chip featured a key structure in which a nanoporous membrane was sandwiched by a cell culture chamber array layer and a corresponding media reservoir array layer. The microfluidic approach took advantage of the characteristics of nanoporous membrane. On one side, this membrane permitted the flow of air but not liquid, thus acting as a flow-stop valve to enable automatic cell distribution. On the other side, it allowed diffusion-based media exchange and thus, mimicked the endothelial layer. In synergy with a liquid transferring platform, the open-access microfluidic system enabled complex multi-step operations involving medium exchange, drug treatment, and cell viability testing. By using this microfluidic protocol, a 10 × 10 tissue arrays was constructed in 90 s, followed by schedule-dependent drug testing. Morphological and immunohistochemical assays results indicated that the resultant tumor tissue was faithful to that in vivo. Drug testing assays showed that the microfluidic tissue array promised multi-step cell assays under biomimetic microenvironment, thus providing an advantageous tool for cell research.