Publicação
Microfluidic-based isolation of rare-tumor associated cells in prostate cancer
| Resumo: | Prostate cancer (PCa) is the most common neoplasia among men. About 90 000 European man die each year from metastatic disease that targets primarily the bones, highlighting the unmet need for new diagnostic tools and therapies. Liquid biopsy has recently been exploited as a promising non-invasive tool towards the molecular profiling of circulating cancer associated cells in various biological fluids, envisaging early diagnosis and real-time patient monitoring of cancer. Among those cells, circulating tumour cells (CTCs), released from the primary tumour into the bloodstream, are considered the main promoters of metastasis. Efficient technologies able to isolate and detect all CTC subpopulations, particularly those most capable of establishing overt metastasis, are urgently required. Other atypical cell population identified in peripheral blood of PCa patients is the cancer-associated macrophages-like cells (CAMLs) that have been reported to interact with CTCs, hitting a role for CAMLs in cancer cell dissemination. Remarkably, CAMLs are more prevalent than CTCs, responsive to treatment and may detect cancer while absent in healthy subjects. In this study, microfluidic cell capture devices have been designed and fabricated for optimized isolation of both CTCs and CAMLs from peripheral blood of PCa patients. Isolation is based on the particular physical characteristics of both target cell populations that allow independent cell collection, in situ molecular characterization and further in vitro culturing. For optimization purposes, monocytes-derived giant cells were firstly generated in vitro, combined with prostate cancer cell line, and spiked in healthy donor blood to be then run into the microfluidic device at different flow rates. Efficiencies of 87% and 43% were obtained in the isolation of PCa cells and monocytes-derived giant cells, respectively (with the flow rate of 100 µL/min for the working fluid and 1000 µL/min for the buffer). Purity of 71% was also achieved. Further experiments with clinical samples from PCa patients are going to be performed in an optimized microfluidic device prototype. The collected two independent cell populations will be then phenotypically characterized in order to find the appropriate combination of molecular markers targeting CTCs and CAMLs and also elucidate about novel molecular targets for future therapeutic approaches. |
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| Autores principais: | Cacho, Ana Rita Pacheco |
| Assunto: | Biomedical microdevices CAMLs CTCs Metastasis Microfluidics Prostate cancer Cancro da próstata Metastização Microdispositivo biomédico Microfluídica |
| Ano: | 2021 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade do Minho |
| Idioma: | inglês |
| Origem: | RepositóriUM - Universidade do Minho |
| Resumo: | Prostate cancer (PCa) is the most common neoplasia among men. About 90 000 European man die each year from metastatic disease that targets primarily the bones, highlighting the unmet need for new diagnostic tools and therapies. Liquid biopsy has recently been exploited as a promising non-invasive tool towards the molecular profiling of circulating cancer associated cells in various biological fluids, envisaging early diagnosis and real-time patient monitoring of cancer. Among those cells, circulating tumour cells (CTCs), released from the primary tumour into the bloodstream, are considered the main promoters of metastasis. Efficient technologies able to isolate and detect all CTC subpopulations, particularly those most capable of establishing overt metastasis, are urgently required. Other atypical cell population identified in peripheral blood of PCa patients is the cancer-associated macrophages-like cells (CAMLs) that have been reported to interact with CTCs, hitting a role for CAMLs in cancer cell dissemination. Remarkably, CAMLs are more prevalent than CTCs, responsive to treatment and may detect cancer while absent in healthy subjects. In this study, microfluidic cell capture devices have been designed and fabricated for optimized isolation of both CTCs and CAMLs from peripheral blood of PCa patients. Isolation is based on the particular physical characteristics of both target cell populations that allow independent cell collection, in situ molecular characterization and further in vitro culturing. For optimization purposes, monocytes-derived giant cells were firstly generated in vitro, combined with prostate cancer cell line, and spiked in healthy donor blood to be then run into the microfluidic device at different flow rates. Efficiencies of 87% and 43% were obtained in the isolation of PCa cells and monocytes-derived giant cells, respectively (with the flow rate of 100 µL/min for the working fluid and 1000 µL/min for the buffer). Purity of 71% was also achieved. Further experiments with clinical samples from PCa patients are going to be performed in an optimized microfluidic device prototype. The collected two independent cell populations will be then phenotypically characterized in order to find the appropriate combination of molecular markers targeting CTCs and CAMLs and also elucidate about novel molecular targets for future therapeutic approaches. |
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