Aarab Meryem, Saadi Salma, Tabyaoui Imane, Badre Latifa, Jouti Nadia Tahiri
Laboratory of Cellular and Molecular Inflammatory, Degenerative, and Oncologic Pathophysiology, Faculty of Medicine and Pharmacy, Hassan II University, Casablanca, Morocco.
Contemp Oncol (Pozn). 2023;27(1):35-40. doi: 10.5114/wo.2023.127190. Epub 2023 Apr 27.
Conventional tissue biopsy is a key examination in cancer diagnosis. However, liquid biopsy is an alternative and less invasive solution that allows the detection of circulating tumour cells (CTCs). CTCs have emerged as a potential screening, diagnostic, and prognostic tool in cancer management. There are many technologies available for the detection and characterization of these cells, but most are either expensive or complicated to apply routinely. Cytological cell blocks (cytoblocks) may be a more practical and cost-effective method to enrich and characterize CTCs and even perform molecular studies. These cytoblocks allow the processing, analysis, and storage of cell suspensions and fluid aspiration samples containing CTCs.
Here we detail a manual protocol based on isolation by density gradient centrifugation, formalin fixing, and paraffin embedding as well as morphological identification for cytological analysis and phenotyping by immunocytochemistry. This method is the result of technical adjustments of previously established protocols.
We succeeded in modifying a protocol for the construction of cytoblocks and applied it to study CTCs in lung and colorectal cancers, respectively.
This less expensive protocol offers a possibility for use in routine diagnosis and can be applied in other fields of research, such as hematology for hematological malignancies and immunology.
传统组织活检是癌症诊断中的关键检查。然而,液体活检是一种替代性的、侵入性较小的解决方案,可用于检测循环肿瘤细胞(CTC)。CTC已成为癌症管理中一种潜在的筛查、诊断和预后工具。有许多技术可用于检测和表征这些细胞,但大多数技术要么成本高昂,要么常规应用复杂。细胞学细胞块(细胞块)可能是一种更实用且具成本效益的方法,用于富集和表征CTC,甚至进行分子研究。这些细胞块可对含有CTC的细胞悬液和液体抽吸样本进行处理、分析和储存。
在此,我们详细介绍一种基于密度梯度离心分离、福尔马林固定、石蜡包埋以及通过免疫细胞化学进行细胞学分析和表型鉴定的手工方案。该方法是对先前既定方案进行技术调整的结果。
我们成功修改了细胞块构建方案,并分别将其应用于研究肺癌和结直肠癌中的CTC。
这种成本较低的方案为常规诊断提供了应用可能性,并且可应用于其他研究领域,如血液学中的血液系统恶性肿瘤研究和免疫学研究。