Journal: Disease Models & Mechanisms

Article Title: Compound design of a patient-derived 3D cell culture system modelling early peritoneal endometriosis

doi: 10.1242/dmm.052436

Figure Lengend Snippet: Characterisation of primary human peritoneal mesothelial cells (HPMCs) and human peritoneal fibroblasts (HPFs) compared to the LP-9 mesothelial cell line and normal human dermal fibroblasts (NHDFs). (A) Representative phase-contrast micrographs of LP-9, HPMCs, NHDFs and HPFs. Mesothelial and fibroblast cells exhibit cobblestone and spindle-like morphologies, respectively. (B) We observed localised expression of the cytoskeletal markers cytokeratin (CK) and vimentin (VIM) in LP-9 cells and HPMCs, while VIM, but not CK, was expressed in NHDFs and HPFs. HPMCs ( n =6) showed a high percentage of CK + /VIM + cells [98.20±1.05% (mean±s.d.)], while HPFs ( n =3) exhibited 81.36±5.63% CK − /VIM + cells. Scale bars: 200 µm (A); 50 µm (B).

Article Snippet: NHDFs were cultured in Fibroblast Growth Medium (Promocell, C-23110) containing 1 ng/ml recombinant human basic fibroblast growth factor and 5 μg/ml recombinant human insulin, supplemented with 2 mM L-glutamine and 100 μg/ml Primocin.

Techniques: Expressing

Journal: Disease Models & Mechanisms

Article Title: Compound design of a patient-derived 3D cell culture system modelling early peritoneal endometriosis

doi: 10.1242/dmm.052436

Figure Lengend Snippet: Establishing composite 3D hydrogel constructs using peritoneal mesothelial cells and fibroblasts. (A) Schematic illustration of model construction and culture timeline. (B) Representative axial view [also seen in C (M3)] and Haematoxylin and Eosin (H&E)-stained section of a hydrogel construct showing the formation of a mesothelial monolayer (ML) and submesothelial layer (SML) on a transwell membrane (TM). (C) Representative images of hydrogel matrices using M1 (collagen I), M2 (70:30 collagen I:Matrigel ratio), M3 (50:50 collagen I:Matrigel ratio) and M4 (collagen I+human fibronectin). Construct generated with matrix combination M3 demonstrated minimal contraction in LP-9/NHDF and HPMC/HPF trials. (D) Lactate dehydrogenase (LDH) cytotoxicity assay in M3 composite hydrogel constructs containing HPMC/HPF ( n =3 donors) over a 10-day culture period. (E) Dual immunofluorescence staining of cleaved caspase-3 (CC-3) and VIM to detect apoptotic HPMCs/HPFs in M3 constructs on day 3 and day 10 of culture ( n =3). Scale bars: 300 µm (B); 100 µm (C); 50 µm (E).

Article Snippet: NHDFs were cultured in Fibroblast Growth Medium (Promocell, C-23110) containing 1 ng/ml recombinant human basic fibroblast growth factor and 5 μg/ml recombinant human insulin, supplemented with 2 mM L-glutamine and 100 μg/ml Primocin.

Techniques: Construct, Staining, Membrane, Generated, LDH Cytotoxicity Assay, Immunofluorescence

Journal: Disease Models & Mechanisms

Article Title: Compound design of a patient-derived 3D cell culture system modelling early peritoneal endometriosis

doi: 10.1242/dmm.052436

Figure Lengend Snippet: Histological and functional analysis of the human parietal peritoneum and peritoneal layer models. (A) Histological staining of transverse sections through parietal peritoneum and composite 3D hydrogel constructs composed of LP-9/NHDFs and HPMCs/HPFs. Immunofluorescence using antibodies against the mesothelial markers podoplanin (PDPN) and mesothelin (MSLN), and submesothelial markers fibroblast specific protein 1 (FSP1) and tumor endothelial marker 1 (TEM1). (B) Colocalisation of MSLN and collagen IV (COLIV) suggesting spontaneous basal lamina formation. (C) Human tissue plasminogen activator (tPA) enzyme-linked immunosorbent assay (ELISA) to determine the functionality of the mesothelial cells in models assembled with HPMCs from three different donors over a 10-day culture period. Scale bars: 50 µm; 15 µm (insets in B).

Article Snippet: NHDFs were cultured in Fibroblast Growth Medium (Promocell, C-23110) containing 1 ng/ml recombinant human basic fibroblast growth factor and 5 μg/ml recombinant human insulin, supplemented with 2 mM L-glutamine and 100 μg/ml Primocin.

Techniques: Functional Assay, Staining, Construct, Immunofluorescence, Marker, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: A host ATPase essential for rhinovirus replication is an antiviral target with a high barrier to resistance

doi: 10.64898/2026.05.13.723454

Figure Lengend Snippet: (A-C) CB-6644 antiviral assays in cell lines. (A) HeLa-H1 cells, (B) BEAS-2B cells or (C) HeLa-E8 cells were infected with the indicated RV types (MOI 20 for A , MOI 1 for B-C ). Cells were treated at 1 hpi with DMSO or the indicated concentrations of CB-6644. Viral titres were quantified at the indicated times post-infection. N=4 or 5 independent experiments. (D-G) CB-6644 antiviral assays in WD-PNECs. (D) Workflow for generation of WD-PNEC cultures. Primary nasal epithelial cells (PNECs) were sampled via nasal brushing from volunteers, expanded in monolayers, and seeded into Transwells. When 100% confluent, after 4-8 days of incubation, apical medium was removed to initiate air-liquid interface (ALI), which triggers cell differentiation and the formation of a pseudostratified epithelium containing ciliated epithelial cells, goblet cells and basal cells. After 28 days of incubation, high quality WD-PNEC cultures were infected apically with the indicated RV (MOI 0.01). CB-6644 or DMSO was added apically 16 h before (E) or at different time points after (F) infection, as indicated. Viral titres in apical washes collected at the indicated times were quantified. N= 3 (E, RV-A16 and RV-B14) or 2 (E, RV-C15 and F) independent donors. (G) Viability of WD-PNECs apically treated with 2 μM CB-6644 or DMSO for 192 h, or with 1% Triton X-100 (TX100) for 2 h, presented as percentage viability relative to DMSO-treated control. N= 3 independent donors. For panels A-C and G, data are shown as individual points, coded by shape according to experimental replicate, with means. For panels E-F, data are shown as means (± SD) connected by lines colour-coded by treatment. Statistical tests: two-tailed paired t -test (A-C), one-way ANOVA with Dunnett’s post-hoc test (G). **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant. See also Figure S2.

Article Snippet: After sampling (5 turns within 1 nostril), nasal brushes were placed inside 15 ml conical tubes and washed with Airway Epithelial Cell Growth Medium (C-21160, Promocell) supplemented with 1X Airway Epithelial Cell Growth Medium supplement pack (C-39160, Promocell) and 1% (v/v) penicillin/streptomycin (15140122, Thermo Fisher Scientific) (monolayer medium).

Techniques: Infection, Incubation, Cell Differentiation, Control, Two Tailed Test