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使用微腔阵列对胃循环肿瘤细胞进行快速回收和短期培养

Rapid Recovery and Short-Term Culture of Gastric Circulating Tumor Cells Using Microcavity Array.

作者信息

Yoshino Tomoko, Takabayashi Tomohiro, Bao Qian, Tanaka Tsuyoshi, Negishi Ryo, Shimoyama Tatsu, Sawada Takeshi, Kanemasa Yusuke, Koizumi Fumiaki

机构信息

Division of Biotechnology and Life Science Institute of Engineering Tokyo University of Agriculture and Technology Koganei Tokyo Japan.

Department of Medical Oncology Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital Tokyo Japan.

出版信息

Eng Life Sci. 2025 Jun 15;25(6):e70026. doi: 10.1002/elsc.70026. eCollection 2025 Jun.

Abstract

Circulating tumor cells (CTCs) hold significant promise for cancer diagnosis, prognosis, and treatment monitoring. We previously developed a technique for a single-cell filtering device known as the microcavity array (MCA), specifically designed for the efficient recovery of CTCs from whole blood samples. Efficient enrichment and release of cells from the MCA remains challenging because of cell adhesion that occurs on the MCA surface during the enrichment phase. This study investigated the effects of surface modification with 2-methacryloyloxyethyl phosphorylcholine (MPC) on the recovery efficiency of cancer cell lines from MCA. Scanning electron microscope (SEM) demonstrated reduced cell-substrate interactions, leading to improved recovery efficiency. Comparative analyses showed that the MCA method provided superior recovery efficiency and reduced processing time compared to traditional methods such as density gradient centrifugation (DGC), while maintaining cell viability and proliferative capacity. CTCs were successfully detected in patients with gastric cancer, and short-term cultures were achieved even when fewer than 20 CTCs per milliliter of blood were isolated. These findings emphasize the importance of surface modification for enhancing CTC isolation and the need for optimized culture conditions. The optimized MCA method offers a promising approach for rapid CTC recovery and potential integration with automated systems. : The Microcavity array (MCA) is a device specifically designed for efficient recovery of CTCs from whole blood. However cell adhesion on the MCA surface can limit release efficiency. This study demonstrated that surface modification with MPC signigicantly reduces cell-substrate adhesion, improving recovery efficiency while maintaining cell viability and proliferative capacity. Compared to traditional density gradient centrifugation, the MPC-modified MCA offers shorter processing time and better performance. CTCs were successfully detected in gastric cancer, and short-term cultures were achieved even when fewer than 20 CTCs per mL of blood were isolated. The method supports downstearm applications such as cancer cell characterization and treatment monitoring. With potential for integration into automated system, the optimized MCA provides a practical, scalable solution for clinical liquid biopsy and personalized oncology.

摘要

循环肿瘤细胞(CTCs)在癌症诊断、预后评估和治疗监测方面具有巨大潜力。我们之前开发了一种用于单细胞过滤装置的技术,即微腔阵列(MCA),它是专门为从全血样本中高效回收CTCs而设计的。由于在富集阶段细胞会粘附在MCA表面,因此从MCA中高效富集和释放细胞仍然具有挑战性。本研究调查了用2-甲基丙烯酰氧乙基磷酰胆碱(MPC)进行表面修饰对从MCA中回收癌细胞系效率的影响。扫描电子显微镜(SEM)显示细胞与底物的相互作用减少,从而提高了回收效率。对比分析表明,与密度梯度离心(DGC)等传统方法相比,MCA方法具有更高的回收效率和更短的处理时间,同时能保持细胞活力和增殖能力。在胃癌患者中成功检测到了CTCs,即使每毫升血液中分离出的CTCs少于20个,也实现了短期培养。这些发现强调了表面修饰对增强CTCs分离的重要性以及优化培养条件的必要性。优化后的MCA方法为快速回收CTCs以及与自动化系统的潜在整合提供了一种有前景的方法。:微腔阵列(MCA)是一种专门为从全血中高效回收CTCs而设计的装置。然而,MCA表面的细胞粘附会限制释放效率。本研究表明,用MPC进行表面修饰可显著降低细胞与底物的粘附,提高回收效率,同时保持细胞活力和增殖能力。与传统的密度梯度离心相比,MPC修饰的MCA处理时间更短,性能更好。在胃癌中成功检测到了CTCs,即使每毫升血液中分离出的CTCs少于20个,也实现了短期培养。该方法支持下游应用,如癌细胞表征和治疗监测。由于有可能集成到自动化系统中,优化后的MCA为临床液体活检和个性化肿瘤学提供了一种实用、可扩展的解决方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f78b/12167759/630a4e31e293/ELSC-25-e70026-g001.jpg

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