Journal: Scientific Reports

Article Title: An exploratory in vitro co-culture of enteric neurons and smooth muscle cells demonstrates neuronal contribution to muscle layer formation

doi: 10.1038/s41598-026-39409-3

Figure Lengend Snippet: Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells). SMCs alone, with direct contact and without direct contact to isolated myenteric plexus cells.

Article Snippet: Fig. 8 Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells).

Techniques: Co-Culture Assay, Isolation

Journal: Scientific Reports

Article Title: An exploratory in vitro co-culture of enteric neurons and smooth muscle cells demonstrates neuronal contribution to muscle layer formation

doi: 10.1038/s41598-026-39409-3

Figure Lengend Snippet: Images of 3 weeks old co-culture of ENS cells and muscle cells in different layouts: ( a ) SMCs alone, ( b ) paracrine interaction between SMCs and ENS cells, ( c ) direct contact between SMCs and ENS cells Scale bars 100 μm, ( d ) paracrine interaction between SMCs and ENS in higher magnification, ( e ) Direct contact between smooth muscle and ENS cells in higher magnification.– muscle cells Scale bars 20 μm, ( f ) 3D reconstruction of muscle fibers in confocal microscopy by the direct contact between SMCs and ENS, ( j ) neurons, which are intercommunicated in the neuronal net within the muscle layer by the direct contact between SMCs and ENS. Confocal microscopy, Green (ß-Tubulin III) – neurons, red (smooth muscle actin (SMA)) – muscle cells. Scale bars 50 μm.

Article Snippet: Fig. 8 Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells).

Techniques: Co-Culture Assay, Confocal Microscopy

Journal: Scientific Reports

Article Title: An exploratory in vitro co-culture of enteric neurons and smooth muscle cells demonstrates neuronal contribution to muscle layer formation

doi: 10.1038/s41598-026-39409-3

Figure Lengend Snippet: Imaging of the whole thickness of the 3D scaffolds after 3 weeks of ENS cells and muscle cells co-culture in different layouts: ( a ) SMCs without ENS cells, ( b ) paracrine interaction between SMCs and ENS cells, ( c ) direct contact between SMCs and ENS cells. Confocal microscopy, Green (ß-Tubulin III) – neurons, red (smooth muscle actin (SMA)) – muscle cells.

Article Snippet: Fig. 8 Schematic representation of plates with inserts and different layouts of muscle and ENS cells in a three-dimensional matrix: ( a ) plates with insert, which allows the medium to surround and support the co-culture from all sides, ( b ) SMCs without ENS cells in the 3D-matrix, ( c ) SMCs and ENS cells equally distributed in 3D-matrix, ( d ) SMCs distributed in upper and lower layers, ENS cells - in the middle layer of the 3D-matrix. ( e ) SMCs distributed in upper and lower layers, ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix. ( f ) SMCs and ENS cells distributed in the middle layer of the 3D-matrix. ( g ) mixed SMCs and ENS cells densely arranged in the same plane of the middle layer of the 3D-matrix, ( h ) different layouts between SMCs and isolated myenteric plexus (ENS cells).

Techniques: Imaging, Co-Culture Assay, Confocal Microscopy

Journal: The Journal of Cell Biology

Article Title: Endothelial Msx1 transduces hemodynamic changes into an arteriogenic remodeling response

doi: 10.1083/jcb.201502003

Figure Lengend Snippet: LSS-dependent canonical BMP signaling triggers MSX1 expression in ECs. (A) HUAECs were transfected with control siRNA or siRNA against SMAD1/5 , seeded in fibronectin-coated chambers, and subsequently kept static or subjected for 1 h to an LSS of 20 dynes/cm 2 . MSX1 expression was analyzed and represented relative to the control static condition. n = 7. (B) Corresponding representative Western blot analysis for MSX1, pSMAD1/5/8, SMAD1, and GAPDH. (C) HUAECs were pretreated for 1 h with DMSO or 2.5-µM DMH1 and subjected to static or LSS (20 dynes/cm 2 ) conditions for 1 h in the presence of DMSO or DMH1. MSX1 expression was analyzed and represented relative to the DMSO condition. n = 3. (D–F) Cells transfected with control siRNA or siRNA against ALK6 ( n = 5), BMPR2 ( n = 6), or ENG ( n = 4) were exposed to static or flow conditions and were analyzed for MSX1 expression. *, P < 0.05 versus the indicated condition. (G) qRT-PCR analysis of HUAECs kept in static conditions or exposed to LSS (20 dynes/cm 2 for the indicated times). mRNA expression is represented relative to the static condition. n = 3. (A, C, and D–F) *, P < 0.05 versus static condition. ud, undetectable. (H) qRT-PCR analysis of HUAECs and SMCs serum starved for 1 h and exposed to 10 ng/µl BMP2, BMP6, or the corresponding amount of solute for 48 h. mRNA expression is represented relative to the control condition. n = 4 and 7. *, P < 0.05; #, P = 0.13 versus corresponding solute condition. Error bars represent the SEM.

Article Snippet: Freshly isolated HUAECs and uterine SMCs (Lonza) were seeded in 6-well plates at a density of 40,000 and 20,000 cells per well, respectively.

Techniques: Expressing, Transfection, Western Blot, Quantitative RT-PCR

Journal: The Journal of Cell Biology

Article Title: Endothelial Msx1 transduces hemodynamic changes into an arteriogenic remodeling response

doi: 10.1083/jcb.201502003

Figure Lengend Snippet: MSX1 induces an inflammatory proadhesive endothelial state. (A) Pictures of THP1 cells (in green) attached to HUAECs upon transduction with control Cherry or MSX1 plasmid for 3 d. Bars, 100 µm. (B) Quantification is represented as the number of monocytes per field of view. n = 6. *, P < 0.05 versus the Cherry control condition. (C) qRT-PCR expression analysis of ICAM1 and VCAM1 in MSX1 overexpression conditions mediated by plasmid transduction of HUAECs ( n = 10) or SMCs ( n = 3). *, P < 0.05 versus the Cherry control condition. (D) qRT-PCR analysis upon siRNA-mediated MSX1 knockdown. mRNA expression is represented relative to the control condition. n = 5–7. *, P < 0.05 versus the corresponding control condition. (E and F) qRT-PCR analysis on HUAECs transfected with control siRNA or siRNA against MSX1 seeded in fibronectin-coated chambers and subsequently kept under static conditions or subjected to 6 h of oscillatory flow. ICAM1 and VCAM1 expression is represented relative to the control static condition. n = 4 and n = 3, respectively. *, P < 0.05 versus the indicated condition. Error bars represent the SEM.

Article Snippet: Freshly isolated HUAECs and uterine SMCs (Lonza) were seeded in 6-well plates at a density of 40,000 and 20,000 cells per well, respectively.

Techniques: Transduction, Plasmid Preparation, Quantitative RT-PCR, Expressing, Over Expression, Transfection

Journal: Diabetologia

Article Title: Native incretins prevent the development of atherosclerotic lesions in apolipoprotein E knockout mice

doi: 10.1007/s00125-011-2241-2

Figure Lengend Snippet: Expression and abundance of GLP-1R and GIPR in monocytes/macrophages and arterial SMCs. a Glp1r and ( b ) Gipr mRNA in exudate peritoneal macrophages and tissues as labelled from non-treated Apoe −/− mice were measured by real-time RT-PCR. c Western blotting analyses of GLP-1R and ( d ) GIPR abundance in exudate peritoneal macrophages of non-treated Apoe −/− mice and other mouse tissues as labelled. Three independent experiments were performed. e Immunostaining of GLP-1R or GIPR in exudate peritoneal macrophages from non-treated Apoe −/− mice. Green, GLP-1R or GIPR as indicated; red, nuclei; red + green, overlay of GLP-1R or GIPR and nuclei. Scale bars 50 μm. f GLP1R and ( g ) GIPR mRNA levels in human coronary artery SMCs, THP1 cells and THP1-derived macrophages. h GIPR mRNA level in human monocytes and monocyte-derived macrophages were measured by real-time RT-PCR. Results were obtained from three to six independent experiments

Article Snippet: The expression of GIPR mRNA in human monocytes was far higher than in human macrophages and human coronary artery SMCs (Lonza) (Fig. ).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunostaining, Derivative Assay