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: Redox Biology

Article Title: Modification of the dermal matrix by senescence associated lipids and its functional consequence

doi: 10.1016/j.redox.2026.104069

Figure Lengend Snippet: Keratinocytes cultured on modified collagen type IV show oxidative stress and reduced MMP levels. A Scatter plots show gene expression levels of IL1A and HMOX1 relative to B2M in keratinocytes cultured on HNE or OxPAPC modified collagen type IV. Primary keratinocytes from two donors (f38y, m27y) were grown in quadruplicates for up to 72 h, n = 8. B Heatmap of log 2 -transformed relative expression levels normalized to the corresponding control. Transcriptional levels of senescence, inflammation, and keratinocyte differentiation markers were assessed. Colors represent expression changes (blue = downregulation, red = upregulation). Statistical significance was determined by one-way ANOVA and is indicated by asterisks (* p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.001) with p-values shown for z values > 1.

Article Snippet: After gelation at 37 °C in a humidified atmosphere for 3 h in the absence of CO 2 , gels were equilibrated in Keratinocyte Growth Medium (KGM2, PromoCell, Germany) and placed into an incubator with 5% CO 2 .

Techniques: Cell Culture, Modification, Gene Expression, Transformation Assay, Expressing, Control

Journal: Redox Biology

Article Title: Modification of the dermal matrix by senescence associated lipids and its functional consequence

doi: 10.1016/j.redox.2026.104069

Figure Lengend Snippet: Skin equivalents with modified collagen show disturbed differentiation and early senescence. A Hematoxylin & Eosin staining of skin equivalents. Collagen was modified, fibroblasts from different donors (f34y, f42y, m25y) were seeded into the matrix, and keratinocytes (f49y, f35y, f30y) on top; n = 6. SE were cultured at the air-liquid interface for 5 days to allow differentiation. B Quantification of nuclei in stratum corneum, indicating parakeratosis, n = 10. Bar graph shows mean ± SD; significant differences determined by one-way ANOVA (* p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.001). C Representative images of SE stained with anti -KRT10, anti -KRT14, and Hoechst nuclear counterstaining. D Boxplot showing quantification of epidermal thickness, n = 4. Data are represented as median ± interquartile range; significance was determined by one-way ANOVA. E Protein expression of cell cycle inhibition marker P16 and housekeeping marker TUBULIN from the epidermis of the SE. F Bar chart showing quantification of P16 vol intensity normalized to TUBULIN with mean ± SD, n = 4, differences determined by one-way ANOVA. G Scatter plots showing expression levels of LMNB1, CDKN1A , and HSPA1A relative to B2M from epidermis and dermis of SE, n = 6. Asterisks represent significant differences determined by one-way ANOVA. H Confocal images of SE with anti -LMNB1 immunostaining and Hoechst nuclear counterstaining. I Boxplot showing quantification of LaminB1 mean signal intensity per pixel, measured in a virtual cross section through the epidermal cells, n = 4, significance determined by one-way ANOVA. J Confocal images of SE with anti -γH2AX immunostaining and Hoechst counterstaining. K Bar chart showing quantification of γH2AX positive nuclei by tissue cytometry analysis of the entire tissue section, mean ± SD, n = 3. Significant differences (* p < 0.05; ** p < 0.01) determined by Student's t - test. Scale bars: A = 50 μm; C = 100 μm; H, J = 20 μm.

Article Snippet: After gelation at 37 °C in a humidified atmosphere for 3 h in the absence of CO 2 , gels were equilibrated in Keratinocyte Growth Medium (KGM2, PromoCell, Germany) and placed into an incubator with 5% CO 2 .

Techniques: Modification, Staining, Cell Culture, Expressing, Inhibition, Marker, Immunostaining, Cytometry

Journal: Journal of Extracellular Biology

Article Title: Proteomic Epithelial‐To‐Mesenchymal Transition Signature in Fetoplacental Small Extracellular Vesicles of Early‐Onset Preeclampsia

doi: 10.1002/jex2.70122

Figure Lengend Snippet: Characterisation of sEVs secreted by fetoplacental endothelial cells (fpECs) from term (T, n = 6), preterm (PT, n = 6) and Early‐Onset preeclamptic (EO‐PE, n = 4) pregnancies. (A) Representative transmission electron microscopy (TEM) images of sEVs (scale bar: 200 nm). (B) NTA: Size and concentration of particles, with size (x‐axis) plotted against particle concentration normalised to cell count at the time of isolation (y‐axis). Data are presented as mean ± standard error (SE). (C) Violin plots display the total concentration of sEVs (normalised to cell count at isolation) as measured by NTA. Statistical analysis was performed using the Kruskal–Wallis test, followed by pairwise Wilcoxon rank‐sum tests with Bonferroni correction for multiple comparisons. Adjusted p values are indicated (* = p < 0.05; ns = not significant). (D) Western blot analysis of sEVs using established sEV markers (Alix, TSG101, Syntenin‐1), endothelial marker (CD31) and non‐vesicular marker (ApoB).

Article Snippet: The cell pellet was resuspended in Endothelial Cell Growth Medium MV (ECGM; PromoCell, C‐22220, Heidelberg, Germany), supplemented with endothelial cell growth factor, hydrocortisone, epidermal growth factor (PromoCell, C‐39220), 0.05 mg/mL gentamicin (Gibco) and 10% defined FCS and plated in 12‐well plates coated with 1% gelatine (Sigma–Aldrich, St. Louis, MO).

Techniques: Transmission Assay, Electron Microscopy, Concentration Assay, Cell Characterization, Isolation, Western Blot, Marker

Journal: Journal of Extracellular Biology

Article Title: Proteomic Epithelial‐To‐Mesenchymal Transition Signature in Fetoplacental Small Extracellular Vesicles of Early‐Onset Preeclampsia

doi: 10.1002/jex2.70122

Figure Lengend Snippet: Descriptive statistics of proteomic analysis of fetoplacental‐endothelial derived sEVs from T ( n = 4), PT ( n = 5) and EO‐PE ( n = 4) pregnancies. (A) Venn diagram of identified proteins in the respective groups. Proteins were filtered to include those with >75% valid values in each group, resulting in 1098 proteins identified in T‐sEVs, 1179 proteins in PT‐sEVs and 1045 proteins in EO‐PE‐sEVs. (B) Principal component analysis (PCA) of protein abundances in the proteome. PCA revealed distinct clustering between the proteomes of EO‐PE‐ and PT‐sEVs, while T‐sEVs overlapped both, indicating shared variability. (C) Volcano plots of quantified proteins. Significant proteins are highlighted in dark grey (T), blue (PT) and red (EO‐PE) circles (two‐sided t ‐test with p values corrected for FDR, statistically significant if absolute log 2 FC ≥ 1.2 and p value ≤ 0.05), while non‐significant proteins are depicted in smaller grey circles.

Article Snippet: The cell pellet was resuspended in Endothelial Cell Growth Medium MV (ECGM; PromoCell, C‐22220, Heidelberg, Germany), supplemented with endothelial cell growth factor, hydrocortisone, epidermal growth factor (PromoCell, C‐39220), 0.05 mg/mL gentamicin (Gibco) and 10% defined FCS and plated in 12‐well plates coated with 1% gelatine (Sigma–Aldrich, St. Louis, MO).

Techniques: Derivative Assay