Krakowski Sophia, Campos Sara, Wolff Henri, Bondzio Gabi, Hehnen Felix, Lommel Michael, Kertzscher Ulrich, Geus Paul Friedrich
Deutsches Herzzentrum der Charité, Institute of Computer-Assisted Cardiovascular Medicine, Augustenburger Platz 1, 13353 Berlin, Germany.
Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Charitéplatz 1, 10117 Berlin, Germany.
Diagnostics (Basel). 2025 Jul 26;15(15):1876. doi: 10.3390/diagnostics15151876.
: Liquid biopsy is a minimally invasive alternative to tissue biopsy and is used to obtain information about a disease from a blood sample or other body fluids. In the context of cancer, circulating tumor cells (CTC) can be used as biomarkers to determine the nature of the tumor, its stage of progression, and the efficiency of the administered therapy through monitoring. However, the low concentration of CTCs in blood (1-10 cells/mL) is a challenge for their isolation. Therefore, a minimally invasive medical device (BMProbe™) was developed that isolates CTCs via antigen-antibody binding directly from the bloodstream. Current investigations focus on the process of detaching bound cells from the BMProbe™ surface for cell cultivation and subsequent drug testing to enable personalized therapy planning. : This article presents two approaches for detaching LNCaP cells from anti-EpCAM coated BMProbes™: enzymatic detachment using TrypLE™ and detachment through enzymatic pretreatment with supplementary flow-induced shear stress. The additional shear stress is intended to increase the detachment efficiency. To determine the flow rate required to gently detach the cells, a computational fluid dynamics (CFD) simulation was carried out. : The experimental test results demonstrate that 91% of the bound cells can be detached enzymatically within 10 min. Based on the simulation, a maximum flow rate of 47.76 mL/min was defined in the flow detachment system, causing an average shear stress of 8.4 Pa at the probe edges. The additional flow treatment did not increase the CTC detachment efficiency. : It is feasible that the detachment efficiency can be further increased by a longer enzymatic incubation time or higher shear stress. The influence on the integrity and viability of cells must, however, be considered.
液体活检是一种微创的组织活检替代方法，用于从血液样本或其他体液中获取有关疾病的信息。在癌症背景下，循环肿瘤细胞（CTC）可作为生物标志物，通过监测来确定肿瘤的性质、进展阶段以及所施用治疗的效果。然而，血液中CTC的低浓度（1 - 10个细胞/毫升）对其分离来说是一项挑战。因此，开发了一种微创医疗设备（BMProbe™），它通过抗原 - 抗体结合直接从血流中分离CTC。目前的研究集中在从BMProbe™表面分离结合细胞以进行细胞培养和后续药物测试的过程，从而实现个性化治疗方案规划。 本文介绍了两种从抗EpCAM包被的BMProbes™上分离LNCaP细胞的方法：使用TrypLE™进行酶解分离以及通过补充流动诱导剪切应力进行酶预处理后分离。额外的剪切应力旨在提高分离效率。为了确定温和分离细胞所需的流速，进行了计算流体动力学（CFD）模拟。 实验测试结果表明，91%的结合细胞可在10分钟内通过酶解分离。基于模拟，在流动分离系统中定义了最大流速为47.76毫升/分钟，在探针边缘处产生的平均剪切应力为8.4帕斯卡。额外的流动处理并未提高CTC的分离效率。 通过延长酶孵育时间或增加剪切应力进一步提高分离效率是可行的。然而，必须考虑对细胞完整性和活力的影响。
