Circulating tumor cells (CTCs) serve as crucial metastatic precursor cells, but their study in animal models has been hindered by their low numbers. To address this challenge, we present DanioCTC, an innovative xenograft workflow that overcomes the scarcity of patient-derived CTCs in animal models. By combining diagnostic leukapheresis (DLA), the Parsortix microfluidic system, flow cytometry, and the CellCelector setup, DanioCTC effectively enriches and isolates CTCs from metastatic breast cancer (MBC) patients for injection into zebrafish embryos. Validation experiments confirmed that MDA-MB-231 cells, transplanted following the standard protocol, localized frequently in the head and blood-forming regions of the zebrafish host. Notably, when MDA-MB-231 cells spiked (i.e., supplemented) into DLA aliquots were processed using DanioCTC, the cell dissemination patterns remained consistent. Successful xenografting of CTCs from a MBC patient revealed their primary localization in the head and trunk regions of zebrafish embryos. DanioCTC represents a major step forward in the endeavors to study the dissemination of individual and rare patient-derived CTCs, thereby enhancing our understanding of metastatic breast cancer biology and facilitating the development of targeted interventions in MBC. Summary statement: DanioCTC is a novel workflow to inject patient-derived CTCs into zebrafish, enabling studies of the capacity of these rare tumor cells to induce metastases.
循环肿瘤细胞（CTCs）是关键的转移前体细胞，但其在动物模型中的研究一直受限于数量稀少。为应对这一挑战，我们开发了DanioCTC——一种创新的异种移植工作流程，旨在克服动物模型中患者来源CTCs稀缺的问题。该技术通过整合诊断性白细胞分离术（DLA）、Parsortix微流控系统、流式细胞术及CellCelector平台，成功从转移性乳腺癌（MBC）患者血液中高效富集并分离CTCs，进而将其注射至斑马鱼胚胎。验证实验证实：按标准流程移植的MDA-MB-231细胞在斑马鱼宿主体内主要定位于头部和造血区域。值得注意的是，将掺入DLA样本的MDA-MB-231细胞经DanioCTC流程处理后，其细胞播散模式仍保持稳定。通过对MBC患者CTCs的成功异种移植，发现这些细胞主要定位于斑马鱼胚胎的头部和躯干区域。DanioCTC为研究单个及罕见患者来源CTCs的播散能力提供了重要技术突破，不仅深化了对转移性乳腺癌生物学的理解，更有助于推动MBC靶向干预策略的开发。核心声明：DanioCTC是一种将患者来源CTCs注射至斑马鱼的新型工作流程，为探究这类稀有肿瘤细胞的转移诱导能力提供了研究平台。
肿瘤(癌症)患者之家