Desenvolvimento de dispositivo microfluídico para biópsia líquida de células tumorais circulantes
Liquid biopsy represents an innovative and minimally invasive approach to cancer detection and monitoring, enabling the analysis of circulating tumor biomarkers, such as circulating tumor cells (CTCs), directly from biological fluids. Given the limitations of conventional biopsy methods, which often...
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| Tipo de recurso: | tesis de maestría |
| Estado: | Versión publicada |
| Fecha de publicación: | 2025 |
| País: | Brasil |
| Institución: | Universidade Estadual Paulista (UNESP) |
| Repositorio: | Repositório Institucional da UNESP |
| Idioma: | portugués |
| OAI Identifier: | oai:repositorio.unesp.br:11449/313866 |
| Acceso en línea: | https://hdl.handle.net/11449/313866 |
| Access Level: | acceso abierto |
| Palabra clave: | Células Tumorais Circulantes Meningioma Glioblastoma Dispositivo Microfluídico Biópsia Líquida |
| Sumario: | Liquid biopsy represents an innovative and minimally invasive approach to cancer detection and monitoring, enabling the analysis of circulating tumor biomarkers, such as circulating tumor cells (CTCs), directly from biological fluids. Given the limitations of conventional biopsy methods, which often involve invasive procedures, high costs, and limited repeatability, this study proposed the development and validation of a microfluidic device functionalized with an anti-EpCAM antibody, capable of isolating and capturing CTCs efficiently, selectively, and viably for further analysis. Device fabrication involved creating molds on silicon wafers via photolithography and replicating the microchannel architecture on PDMS using soft lithography. The process showed high reproducibility, structural integrity, and costeffectiveness. Functionalization with avidin and a biotinylated anti-EpCAM antibody provided specificity for tumor cell capture. Flow experiments revealed that increasing injection speed led to higher capture rates, with significantly improved performance in functionalized devices, underscoring the importance of EpCAM as a selection marker. The cell line used is MCF-7 (breast cancer), as well as primary cell culture expansions of glioblastoma and meningioma, all EpCAM-positive, in addition to CCRF-CEM as a negative control. Primary cultures of glioblastoma and meningioma were successfully obtained from surgical samples and expanded without contamination, confirming the experimental model's viability. The device demonstrated not only high capture efficiency and purity but also the ability to release viable cells post-capture, enabling further downstream analyses such as sequencing and single-cell functional assays. In addition to in vitro validations, the device was evaluated on clinical samples from patients with central nervous system tumors, proving its practical applicability by capturing a promising number of CTCs. These results indicate that the proposed device is a valuable tool for tumor diagnosis, monitoring, and the investigation of tumor heterogeneity. Thus, the developed microfluidic system represents a significant advancement in liquid biopsy technology, particularly for central nervous system cancers and other tumor types. Its combination of structural simplicity, low cost, and functional efficiency supports its potential for clinical translation and contributes meaningfully to early diagnosis and personalized cancer care. |
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