Journal: Oncology Letters

Article Title: Diagnostic difficulties in the differentiation between an ovarian metastatic low‑grade appendiceal mucinous neoplasm and primary ovarian mucinous cancer: A case report and literature review

doi: 10.3892/ol.2024.14633

Figure Lengend Snippet: Right ovary with metastatic low-grade appendiceal mucinous neoplasm. Immunohistochemical staining for (A) caudal type homeobox 2, (B) cytokeratin 20 and (C) paired box gene 8. Magnification, ×100.

Article Snippet: Antigen retrieval by microwave and blocking of endogenous peroxidase activity (by 1% hydrogen peroxide in distillated water for 10 min at room temperature) was conducted before incubation with the following primary antibodies: CK20 (monoclonal mouse anti-human antibody; ready-to-use; cat. no. GA777; DAKO; Agilent Technologies, Inc.), CDX2 (monoclonal mouse anti-human antibody; ready-to-use; cat. no. GA080; DAKO; Agilent Technologies, Inc.) and PAX8 (monoclonal mouse anti-human antibody; ready-to-use; cat. no. 760-4618; Roche Diagnostics) at 4°C overnight.

Techniques: Immunohistochemical staining, Staining

Journal: Oncology Letters

Article Title: Diagnostic difficulties in the differentiation between an ovarian metastatic low‑grade appendiceal mucinous neoplasm and primary ovarian mucinous cancer: A case report and literature review

doi: 10.3892/ol.2024.14633

Figure Lengend Snippet: Expression patterns of the most common immunohistochemistry markers depending on the origin of the tumor.

Article Snippet: Antigen retrieval by microwave and blocking of endogenous peroxidase activity (by 1% hydrogen peroxide in distillated water for 10 min at room temperature) was conducted before incubation with the following primary antibodies: CK20 (monoclonal mouse anti-human antibody; ready-to-use; cat. no. GA777; DAKO; Agilent Technologies, Inc.), CDX2 (monoclonal mouse anti-human antibody; ready-to-use; cat. no. GA080; DAKO; Agilent Technologies, Inc.) and PAX8 (monoclonal mouse anti-human antibody; ready-to-use; cat. no. 760-4618; Roche Diagnostics) at 4°C overnight.

Techniques: Expressing, Immunohistochemistry

Journal: Scientific Reports

Article Title: Multicellular ovarian cancer spheroids: novel 3D model to mimic tumour complexity

doi: 10.1038/s41598-024-73680-6

Figure Lengend Snippet: Microscopic characterisation of the structure of simultaneously seeded mono and co-cultured spheroids. OvCar8 (OC), A2780 (OC) and Detroit 551 (Fibr.) monocellular and OvCar8 and Detroit 551, A2780 and Detroit 551 multi-cellular spheroids were cultured for 96 h as described in Fig. A. ( A ) Spheroids were examined by scanning electron microscopy after 96 h of growth. Scale bar 50 μm. ( B,C ) Prior to cultivation, the cells were stained with fluorescence dyes in order to subsequently assign them to a cell type during cultivation. These spheroids were fixed, cleared and imaged after 96 h of growth using LSM. Red/Cell Tracker Deep Red: ovarian cancer cells; green/CellTracker Green CMFDA: fibroblasts. Scale bar 200 μm. (D, E) Histological and immunohistochemical analysis of spheroid morphology and protein expression of OvCar8 ( D ) and A2780 ( E ) spheroids after 96 h of growth. Ki-67 is a proliferation marker, while PAX8 is marker for ovarian cancer cells and fibroblast activation protein markers fibroblasts, respectively. Scale bar 200 μm.

Article Snippet: For immunohistochemistry, sections were stained with antibodies directed against Ki-67 (dilution 1:100, clone SP6, ThermoFisher Scientific), PAX8 (dilution 1:50, clone MRQ-50, Cell Marque), FAP Fibroblast activation protein (dilution 1:400, clone EPR20021, Abcam).

Techniques: Cell Culture, Electron Microscopy, Staining, Fluorescence, Immunohistochemical staining, Expressing, Marker, Activation Assay

Journal: Scientific Reports

Article Title: Multicellular ovarian cancer spheroids: novel 3D model to mimic tumour complexity

doi: 10.1038/s41598-024-73680-6

Figure Lengend Snippet: Morphological characterisation of sequential seeded co-cultured spheroids. OvCar8 and Detroit 551, A2780 and Detroit 551 co-cultured spheroids were seeded time-shifted and cultured for 96 h as described in Fig. A. ( A,B ) Spheroids were examined by scanning electron microscopy after 96 h of growth. Scale bar 50 μm. ( C,D ) Prior to cultivation, the cells were stained with different fluorescence dyes in order to subsequently assign them to a cell type during cultivation. These spheroids were fixed, cleared and imaged after 96 h of growth using LSM. Red/Cell Tracker Deep Red: ovarian cancer cells; green/CellTracker Green CMFDA: fibroblasts. Scale bar 200 μm. ( E,F ) Histological and immunohistochemical analysis of spheroid morphology and protein expression of spheroids after 96 h of growth. Ki-67 is a proliferation marker, while PAX8 is marker for tumour cells and fibroblast activation protein markers fibroblasts, respectively. Scale bar 200 μm.

Article Snippet: For immunohistochemistry, sections were stained with antibodies directed against Ki-67 (dilution 1:100, clone SP6, ThermoFisher Scientific), PAX8 (dilution 1:50, clone MRQ-50, Cell Marque), FAP Fibroblast activation protein (dilution 1:400, clone EPR20021, Abcam).

Techniques: Cell Culture, Electron Microscopy, Staining, Fluorescence, Immunohistochemical staining, Expressing, Marker, Activation Assay

Journal: Virology Journal

Article Title: Oncolytic activity of a coxsackievirus B3 strain in patient-derived cervical squamous cell carcinoma organoids and synergistic effect with paclitaxel

doi: 10.1186/s12985-024-02502-y

Figure Lengend Snippet: Generation of CSCC Organoids. (A) Schematic illustration of the derivation of CSCC organoids from patient samples. (B) Representative brightfield microscopy images of CSCC organoids post-seeding (exemplified by CSCC-2; see Fig. for more examples). Scale bar, 25 μm. (C) Measurements of the growth diameters of CSCC organoids at days 1, 7, and 11. Horizontal lines and error bars represent the mean diameter ± SEM for five organoids ( n = 5). (D) Immunohistological features of CSCC organoids derived from patients, compared to original tumors (exemplified by CSCC-2, see Fig. for more examples). Representative immunofluorescence images display markers Ki67, p53, PanCK, PAX8, and CAR (all in red), with nuclei counterstained using DAPI (blue) and cell membranes with phalloidin (green). Inset images from the left panels are magnified three-fold in the right panels for enhanced detail. Scale bar, 50 μm. (E) Proportion of immunofluorescence staining-positive cells in primary tumors and organoids ( n = 3). Statistical significance was assessed using two-way ANOVA. T, tumor; O, organoid; ns, not significant; * p < 0.05; *** p < 0.001; **** p < 0.0001; ns, not significant

Article Snippet: For immunofluorescence analysis of uninfected CSCC organoids and original tumors, samples cultured in CellCarrier Ultra microplates (PerkinElmer) were fixed with 4% paraformaldehyde, permeabilized with 3% Triton-X, washed with PBS, blocked with 2% BSA for one hour, and incubated overnight at 4 °C with antibodies against Ki67 (Novus Biologicals, NB110-89717), p53 (Cell Signaling Technology, 2527S), PAX8 (Cell Signaling Technology, 59019S), PanCK (Abcam, ab7753), or CAR (Abcam, ab100811).

Techniques: Microscopy, Derivative Assay, Immunofluorescence, Staining