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O carcinoma seroso de alto grau do ovário (CSAG) é o subtipo mais comum e agressivo do cancro do ovário, associado a elevada recidiva e resistência à quimioterapia. A ascite maligna (AM), frequente nestes doentes, representa uma fonte útil para desenvolver modelos ex vivo. Este estudo teve como objetivo estabelecer um protocolo de avaliação histopatológica de organoides derivados de pacientes (ODPs), cultivados a partir de células de AMs com CSAG. Numa fase inicial, testaram-se linhas celulares em duas condições tridimensionais de cultura, tendo o polyHEMA revelado melhor desempenho. Seguidamente, processaram-se oito amostras de AMs do IPO do Porto, cultivadas em meio específico, com ou sem suplementação por sobrenadante autólogo. Após 30 dias, os ODPs foram fixados e analisados histológica e imunocitoquímica, avaliando-se biomarcadores de diferenciação (MUC1, MUC16 e MSLN), proliferação (Ki67), apoptose (caspase 3 clivada) e quimiorresistência (ALDH1). Os resultados mostraram manutenção parcial dos marcadores de diferenciação, mas redução da proliferação, aumento da apoptose e diminuição de ALDH1, sugerindo perda de subpopulações resistentes. Conclui-se que, apesar de alterações fenotípicas ao longo da cultura, os ODPs derivados de AMs são modelos viáveis para estudo funcional do CSAG, representando uma ferramenta promissora para investigação translacional e desenvolvimento de estratégias terapêuticas personalizadas.
High-grade serous ovarian carcinoma (HGSOC) is the most common and aggressive subtype of ovarian cancer, characterized by high recurrence rates and chemotherapy resistance. Malignant ascites (MA), frequently observed in these patients, is a valuable source for developing ex vivo models. This study aimed to establish a histopathological evaluation protocol for patient-derived organoids (PDOs) cultured from MA cells of HGSOC patients. Initially, cell lines were tested under two three-dimensional culture conditions, with polyHEMA showing the best performance. Subsequently, eight MA samples from the Portuguese Oncology Institute of Porto were processed and cultured in a specific medium, with or without autologous supernatant supplementation. After 30 days, PDOs were fixed and analyzed through histology and immunocytochemistry, assessing differentiation, proliferation, apoptosis, and chemoresistance biomarkers. Results showed partial maintenance of differentiation markers, but decreased proliferation, increased apoptosis, and reduced ALDH1 expression, suggesting the loss of resistant subpopulations. In conclusion, despite phenotypic changes during culture, MA-derived PDOs are viable models for functional studies of HGSOC, representing a promising tool for translational research and the development of personalized therapeutic strategies.
High-grade serous ovarian carcinoma (HGSOC) is the most common and aggressive subtype of ovarian cancer, characterized by high recurrence rates and chemotherapy resistance. Malignant ascites (MA), frequently observed in these patients, is a valuable source for developing ex vivo models. This study aimed to establish a histopathological evaluation protocol for patient-derived organoids (PDOs) cultured from MA cells of HGSOC patients. Initially, cell lines were tested under two three-dimensional culture conditions, with polyHEMA showing the best performance. Subsequently, eight MA samples from the Portuguese Oncology Institute of Porto were processed and cultured in a specific medium, with or without autologous supernatant supplementation. After 30 days, PDOs were fixed and analyzed through histology and immunocytochemistry, assessing differentiation, proliferation, apoptosis, and chemoresistance biomarkers. Results showed partial maintenance of differentiation markers, but decreased proliferation, increased apoptosis, and reduced ALDH1 expression, suggesting the loss of resistant subpopulations. In conclusion, despite phenotypic changes during culture, MA-derived PDOs are viable models for functional studies of HGSOC, representing a promising tool for translational research and the development of personalized therapeutic strategies.
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Cancro do ovário Ascites malignas Quimiorresistênca Organóides derivados de pacientes Imunocitoquímica