Human Ovarian Fibroblasts

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers worldwide, with a five year survival rate below 10%. The high mortality rate of PDAC is primaril... More

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers worldwide, with a five year survival rate below 10%. The high mortality rate of PDAC is primarily due to its aggressive metastatic nature, late diagnosis, and resistance to available therapies. A strategic research direction that demands significantly more attention is the identification of novel tumor markers and the development of tools for rapid, noninvasive diagnosis of this type of cancer. Circulating tumor cells (CTCs) are a promising target for early cancer detection via liquid biopsy, but their clinical application in PDAC is limited due to their rarity and technical challenges in reliable isolation and analysis. CTCs are typically isolated based on physical properties (size, density, deformability, charge, shape, hydrodynamics) or antigenic markers using specific antibodies. Although numerous cancer-specific antibodies have been identified, a marker uniquely distinguishing PDAC has yet to be found. In this work, we present the latest results concerning the development of a microfluidic platform that enables effective immunomagnetic isolation of PDAC cells from a complex biological sample using two potential PDAC markers: Plectin-1 and EpCAM. Here, we developed a low-cost, easy-to-manufacture, and user-friendly 3D-printed microfluidic platform that integrates a micromixer and a separation chamber for rapid cell isolation. Optimization tests showed that Plectin-1 at a concentration of 0.485 μg/mL captured 102 cells/ml with a recovery of 80% ± 13.3%, EpCAM at a concentration of 0.625 μg/mL captured 103 cells/ml with a recovery of 47% ± 13.3%, and the mixture of EpCAM and Plectin-1 at concentrations of 0.3125 μg/mL and 0.242 μg/mL, respectively, achieved a recovery of 91.3% ± 4.4%. To demonstrate the applicability of the method in biological samples, blood with different anticoagulants was tested. The highest detection sensitivity was achieved in the blood sample with citrate: Plectin-1 at 0.485 μg/mL enabled detection at a concentration of 102 cells/ml (47 ± 9% recovery), and EpCAM at 1.25 μg/mL achieved a 92 ± 8% recovery. The entire isolation process from biological samples was performed in less than 1 min. Using this system combined with antibody-functionalized magnetic nanoparticles targeting EpCAM and Plectin-1, we demonstrated the efficient capture of PDAC cells from biological samples, highlighting Plectin-1 as a promising PDAC marker and the potential of immunomagnetic microfluidic separation for improved diagnostics. Less

The invasive and metastatic spread of serous ovarian cancer (SOC) results from the cooperative interactions between cancer and stroma, which include extracellular matrix ... More

The invasive and metastatic spread of serous ovarian cancer (SOC) results from the cooperative interactions between cancer and stroma, which include extracellular matrix (ECM) and cellular components, including cancer-associated fibroblasts (CAFs). Soluble factors secreted by cancer and stromal cells contribute to stroma remodeling through the secretion of ECM proteins, providing a favorable environment for cancer cell dissemination. The peptide endothelin-1 (ET-1), through two G protein-coupled receptors (GPCR), endothelin receptor type A (ETAR) and B (ETBR), acts on both cancer and stromal cells, engaging the protein β-arrestin1 (β-arr1), to bolster SOC progression. However, its role in the regulation of the ECM proteins by ovarian fibroblasts is not understood. This study delves into the role of ET-1 as a regulator of type I collagen (Col1) and fibronectin (FN). Less

Three-dimensional (3D) cellular models of cancer tissue are necessary tools to analyze new anticancer drugs under in vitro conditions. Diagnostics and treatment of ovaria... More

Three-dimensional (3D) cellular models of cancer tissue are necessary tools to analyze new anticancer drugs under in vitro conditions. Diagnostics and treatment of ovarian cancer are major challenges for current medicine. In our report we propose a new three-dimensional (3D) cellular model of ovarian cancer which can mimic a fragment of heterogeneous cancer tissue. We used Lab-on-a-chip technology to create a microfluidic system that allows cellular multilayer to be cultured. Cellular multilayer mimics the structure of two important elements of cancer tissue: flesh and stroma. For this reason, it has an advantage over other in vitro cellular models. We used human ovarian fibroblasts (HOF) and human ovarian cancer cells in our research (A2780). In the first stage of the study, we proved that the presence of non-malignant fibroblasts in co-culture with ovarian cancer cells stimulates the proliferation of cancer cells, which is important in the progression of ovarian cancer. In the next stage of the research, we tested the usefulness of the newly-developed cellular model in the analysis of anticancer drugs and therapies under in vitro conditions. We tested two photosensitizers (PS): free and nanoencapsulated meso-tetrafenylporphyrin, and we evaluated the potential of these drugs in anticancer photodynamic therapy (PDT) of ovarian cancer. We also studied the mechanism of PDT based on the analysis of the level of reactive oxygen species (ROS) in cell cultures. Our research confirmed that the use of new-generation PS can significantly increase the efficacy of PDT in the treatment of ovarian cancer. We also proved that the newly-developed 3D cellular model is suitable for rapid screening of anticancer drugs and has the potential to be used clinically in the future, e.g. in the selection of treatment methods for anticancer personalized medicine. Less