Journal: International Journal of Molecular Sciences

Article Title: Tissue-Engineering the Fibrous Pancreatic Tumour Stroma Capsule in 3D Tumouroids to Demonstrate Paclitaxel Response

doi: 10.3390/ijms22084289

Figure Lengend Snippet: Pancreatic ductal adenocarcinoma and its desmoplastic, hypovascular tumour stroma. This fibrotic capsule contains a high concentration of extracellular matrix (ECM) components such as collagen, hyaluronic acid, laminins, and fibronectin partially produced by a dense population of cancer-associated fibroblasts. This schematic focuses on ECM components within the 3D tumouroid model. Created using Servier Medical Art according to the Creative Commons Attribution 3.0 Unported License guidelines 3.0 ( https://creativecommons.org/licenses/by/3.0/ , accessed on 9 February 2021).

Article Snippet: We tissue-engineered a biomimetic 3D pancreatic cancer (“tumouroid”) model comprised of a central artificial cancer mass (ACM), containing MIA Paca-2 cells, surrounded by a fibrotic stromal compartment.

Techniques: Concentration Assay, Produced

Journal: Development (Cambridge, England)

Article Title: Development of a 3D atlas of the embryonic pancreas for topological and quantitative analysis of heterologous cell interactions

doi: 10.1242/dev.199655

Figure Lengend Snippet: Analysis of pancreatic morphogenesis using confocal microscope and LSFM. (A,A′) Schematics of the pancreatic epithelium, spleno-pancreatic mesenchyme, and developing vasculature in E12.5 and E14.5 mouse embryos in 3D (A) or transverse section (A′). Depiction of tissue architecture in the transverse section highlights the segregation of the pancreatic epithelium into domains of ‘tip’ and ‘trunk’ cells from E12.5 onwards. A, anterior; L, left; P, posterior; R, right. (B) Genetic strategy for labeling distinct mouse pancreatic, endothelial or mesenchymal cell populations. When not recombined, the Tg reporter construct mTmG results in the ubiquitous expression of a membrane-targeted tdTomato protein (mT). Cre-mediated recombination results in the excision of the mT expression cassette and expression of the membrane-targeted GFP (mG) in a tissue-specific fashion . (C,D) IF detection of mG (green) and mT (red) in pancreatic tissue of E12.5 Tg( Pdx1 -Cre, mTmG) embryos (C) or E14.5 Tg( Nkx2.5 -Cre, mTmG) embryos (D). Pancreata analyzed as tissue sections by confocal microscopy (left) or as whole-mount preparations by LSFM (right) are shown side by side. In Pdx1 -Cre Tg embryos (C), mG marks pancreatic epithelial cells. In Nkx2.5 -Cre embryos (D), mG labels a subpopulation of mesenchymal cells in the spleno-pancreatic mesenchyme. Hoechst dye (gray) was used as nuclear counterstain on tissue sections. (E,F) Representative images of IF labeling of E12.5 and E14.5 pancreatic tissue. IF for Ecad (red; E,F) and Pdx1 (green; E) marks the pancreatic epithelium, whereas IF for ERG (blue; F) and VEcad (green; F) identifies endothelial cells surrounding the developing pancreas. Scale bars: 100 μm.

Article Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5.

Techniques: Microscopy, Labeling, Construct, Expressing, Membrane, Confocal Microscopy

Journal: Development (Cambridge, England)

Article Title: Development of a 3D atlas of the embryonic pancreas for topological and quantitative analysis of heterologous cell interactions

doi: 10.1242/dev.199655

Figure Lengend Snippet: 3D visualization of tissue interactions during pancreatic development using LSFM. (A) Schematic of the experimental setup for collection, visualization and imaging of pancreatic tissue from mouse embryos using LSFM. (B-G) Representative LSFM 3D images of pancreatic tissue from Tg mouse embryos at E12.5 (B,D,F) or E14.5 (C,E,G). Lineage-specific Cre induced mG expression in the indicated specific cell types. In Ins2 -Cre; mTmG Tg embryos (B,C), mG marks insulin-expressing cells. In Nkx2.5 -Cre; mTmG Tg embryos (D,E), mG labels a subset of mesenchymal cells in the spleno-pancreatic mesenchyme. In Cdh5 -Cre; mTmG Tg embryos (F,G), mG marks endothelial cells. In addition to detection of mG and mT, samples were stained for Pdx1 (D,E; blue) or Ecad (B,C,F,G; blue) to visualize pancreatic epithelial cells. A, anterior; L, left; P, posterior; R, right. Scale bars: 100 μm.

Article Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5.

Techniques: Imaging, Expressing, Staining

Journal: Development (Cambridge, England)

Article Title: Development of a 3D atlas of the embryonic pancreas for topological and quantitative analysis of heterologous cell interactions

doi: 10.1242/dev.199655

Figure Lengend Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5. WMIF for Ecad (A,B; green) or Pdx1 (C,D; blue) labels the pancreatic epithelium and for VEcad (red; A,B) or ERG (blue; C,D) marks the endothelium. mG (green; C,D) and mT (red; C,D) in Tg( Nkx2.5 -Cre; mTmG) embryos mark the Nkx2.5-Cre + mesenchyme and Nkx2.5-Cre − tissues, respectively. 3D images and surface renderings are shown as merged (left) and individual channels (right). A, anterior; L, left; P, posterior; R, right. Scale bars: 100 μm. (E) Quantification of the mean diameters of blood vessels in close proximity (<15 µm) to the pancreatic epithelium at E12.5 ( n =3) and E14.5 ( n =2). Mann–Whitney test, *** P <0.001. Data are mean±s.d.

Article Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5.

Techniques: MANN-WHITNEY

Journal: Development (Cambridge, England)

Article Title: Development of a 3D atlas of the embryonic pancreas for topological and quantitative analysis of heterologous cell interactions

doi: 10.1242/dev.199655

Figure Lengend Snippet: Quantitative analysis of endothelial and mesenchymal cell distribution around the pancreatic epithelium. (A,B) 3D rendering of LSFM scans of E12.5 and E14.5 pancreas and surrounding tissues labeled with the indicated antibodies. Ecad + (A) or Pdx1 + (B) pancreatic epithelial cells were rendered as surfaces (gray). ERG + endothelial (spots, A) or Nkx2.5-Cre + mesenchymal (spots, B) cells were identified using ‘Spots’ function in Imaris and color-coded based on their distance from the epithelium. Only endothelial or mesenchymal cells within a 15 μm distance from the epithelium were considered in the analysis. A, anterior; L, left; P, posterior; R, right. Scale bars: 200 μm. (C,D) Violin plots showing the distribution of endothelial (C) or Nkx2.5-Cre + mesenchymal (D) cells around the pancreatic epithelium at E12.5 [ n =799 cells (C); n =3350 cells (D)] and E14.5 [ n =7817 cells (C); n =3359 cells (D)]. Mann–Whitney test, *** P <0.001. (E,F) Analysis of endothelial (E) or Nkx2.5-Cre + mesenchymal (F) cell distance from pancreatic epithelium presented as cumulative frequency distribution. The percentage of endothelial (E) or Nkx2.5-Cre + mesenchymal (F) cell population is plotted against the distance from the epithelium. Dashed vertical line indicates the 7.5 µm boundary, which corresponds approximately to the thickness of the first cell layer in direct contact with the epithelium. (G) Representative images of Ecad (orange) and ERG (green) IF labeling in E12.5 pancreas and stomach sections. Hoechst (blue) was used as nuclear counterstain. Scale bars: 50 µm. (H) Scatter plot showing quantification of endothelial cell distance from the pancreas or stomach epithelium at E12.5 ( n =3) and E14.5 ( n =3). Quantification was performed using HALO software. Data represents the number of endothelial cells localized within the first cell layer from the epithelium as a percentage of the total number of endothelial cells within a distance <15 µm from the pancreas or stomach. Kruskal–Wallis test, ** P <0.001. Data are mean±s.d.

Article Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5.

Techniques: Labeling, MANN-WHITNEY, Software

Journal: Development (Cambridge, England)

Article Title: Development of a 3D atlas of the embryonic pancreas for topological and quantitative analysis of heterologous cell interactions

doi: 10.1242/dev.199655

Figure Lengend Snippet: Tissue segmentation of murine embryonic pancreas. (A) 3D representation of LSFM scans of E12.5 and E14.5 pancreas and surrounding tissues labeled with antibodies against Ecad (epithelial cells; green) and ERG (endothelial cells; orange). DRAQ5 was used as nuclear counterstain. The ‘Spot detection’ function in Imaris was used to identify all cells (based on DRAQ5), individual epithelial (Ecad + ) or endothelial cells (ERG + ) in the indicated boxed regions. Mesenchymal cells were identified as DRAQ5 + /Ecad − /ERG − . Analysis was restricted to surrounding tissue within 15 µm distance from the epithelial surface. Right-hand panels show higher magnification single-channel images of the boxed region. Scale bars: 100 µm. (B) Representative image of Ecad and ERG IF labeling of E12.5 pancreatic tissue sections (left). Hoechst was used as nuclear counterstain. Epithelial, endothelial and mesenchymal cells were segmented using HALO software (middle). Right-hand panel shows higher magnification of the boxed region. Mesenchymal cells were identified as Ecad − /ERG − /Hoechst + . Scale bars: 50 µm (left and middle); 20 µm (right). (C,D) Scatter plots of the relative abundance of endothelial, epithelial and mesenchymal cells within a 15 µm radius of pancreatic tissue, shown as percentage of total cell numbers. Analysis of cell type fractions in C was performed using Imaris software on LSFM of whole-mount samples (A), whereas the analysis in D was performed using HALO software on confocal images of IF sections (B). No differences in cell fractions were detected between embryonic stages (E12.5 versus E14.5) or analysis pipelines (Imaris versus HALO). Kruskal–Wallis test, followed by Dunn's multiple comparisons test. Data are mean±s.d.

Article Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5.

Techniques: Labeling, Software

Journal: Development (Cambridge, England)

Article Title: Development of a 3D atlas of the embryonic pancreas for topological and quantitative analysis of heterologous cell interactions

doi: 10.1242/dev.199655

Figure Lengend Snippet: 3D rendering of pancreatic tissue. (A) Representative LSFM 3D images of the endocrine pancreas from wild-type embryos at E14.5, E16.5 and newborns at P0. WMIF for VEcad (red) marks the endothelium, glucagon (blue) labels the endocrine α-cells, Pdx1 (green) labels the pancreatic epithelium at E14.5 and becomes enriched in endocrine cells from E16.5 onward. 3D images are shown as merged (leftmost panels) and individual channels. (B) Surface renderings of the boxed area in A show glucagon + cells forming budding peninsular structures attached to the epithelial cords. Insets display the boxed area at higher magnification. Scale bars: 100 μm.

Article Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5.

Techniques:

Journal: Development (Cambridge, England)

Article Title: Development of a 3D atlas of the embryonic pancreas for topological and quantitative analysis of heterologous cell interactions

doi: 10.1242/dev.199655

Figure Lengend Snippet: Quantitative analysis of endothelial and mesenchymal cell density around the pancreatic endocrine clusters. (A,B) 3D rendering of LSFM scans of E14.5 and E16.5 pancreata from wild-type or Tg( Nkx2.5 -Cre; R26R-H2B-GFP) embryos. WMIF for glucagon (white) labels the endocrine α-cells, VEcad (red) marks the endothelium (A), and Nkx2.5-Cre + mesenchymal cells are GFP + (green) (B). Central and right panels show endocrine clusters composed of at least eight glucagon + cells (3D rendering volume >67,700 µm 3 ) rendered as surfaces (white) for quantitative analyses. Endothelial and mesenchymal cells within the close (up to 15 μm; central panels) and distant (up to 50 μm, right panels) microenvironment from the endocrine clusters were considered in quantitative analyses. Scale bars: 100 μm. (C,D) Vascular structure diameter distribution (relative frequency as percentage) in the microenvironment surrounding the endocrine clusters at E14.5 ( n =3) (C) and E16.5 ( n =3) (D). Data are mean±s.d. (E) Scatter plots showing the vascular volume fraction measurement at a distance of 0-15 µm and 15-50 µm from the endocrine clusters at E14.5 and E16.5. The vascular volume fraction was used as proxy for the vascular density, shown as ratio between the volume of the VEcad + surface and the volume of the analyzed area (0-15 or 15-50 μm away from glucagon + surface). (F) Scatter plots showing the mesenchymal density (Nkx2.5-GFP + cells) in the microenvironment surrounding the endocrine clusters (glucagon+) at E14.5 and E16.5.

Article Snippet: 3D rendering of pancreatic tissue. (A-D) Representative LSFM 3D images (top) and Imaris surface renderings (bottom) of the developing pancreas from wild-type (A,B) or Tg( Nkx2.5 -Cre; mTmG) (C,D) embryos at E12.5 and E14.5.

Techniques:

Journal: Frontiers in Immunology

Article Title: Translational strategy to support the first-in-human study of a TCR-like T cell bispecific with an in vitro -based safety approach

doi: 10.3389/fimmu.2026.1736584

Figure Lengend Snippet: Evaluation of HLA mismatch impact on co-culture sensitivity and background activation. (A) Liver spheroid co-cultures comparing autologous and allogeneic (HLA-A*02 matched) effector donors. Data are presented as mean ± SD as shaded area of technical replicates (n=3) from 3 representative donor pairs (N = 3) for each HLA-A*02 background. Statistical significance relative of the autologous cultures compared to corresponding allogenic cultures two-way ANOVA with Tukey’s post-hoc test, p values are indicated for the overall comparison between autologous versus allogeneic data. (B) Pancreatic islet spheroid co-cultures comparing HLA-matched (4/6 alleles) and HLA-mismatched conditions. Data illustrate the increased signal-to-noise ratio in HLA-matched systems compared to the elevated background activation observed in allogeneic co-cultures. Data are presented as mean of N = 5 donors for the matched dataset and N = 4 donors for the unmatched dataset run in technical replicates (n=2).

Article Snippet: Pancreatic Islets: 3D InSight Islet Microtissues (Cat: MT-04-002-01-60, InSphero) composed of primary alpha, beta, and delta cells were used.

Techniques: Co-Culture Assay, Activation Assay, Comparison

Journal: Journal of Surgical Case Reports

Article Title: Refractory pancreatic fistula following distal pancreatectomy successfully treated with N-butyl-2-cyanoacrylate and vascular embolization coils using a pull-through technique

doi: 10.1093/jscr/rjag358

Figure Lengend Snippet: (a) The micro catheter was guided into the pancreatic duct through the percutaneous drain site over a guidewire. (b) Vascular embolization coils were placed in the pancreatic duct stump, followed by the injection of NBCA mixed with lipiodol into the coil deployment site.

Article Snippet: Vascular embolization coils (Target 3D Detachable Coils, Stryker) were deployed into the pancreatic duct stump, followed by NBCA injection ( and ).

Techniques: Injection