bronchial smooth muscle cells hbsmc hbsmc  (PromoCell)

Journal: PLOS One

Article Title: β-Nicotinamide adenine dinucleotide (β-NAD) acts as a bronchodilator

doi: 10.1371/journal.pone.0334491

Figure Lengend Snippet: (A, B) Force recordings in organ bath, β-NAD (A) and salbutamol (B) concentration-dependently relax precontracted human bronchioli. n refers to the number of bronchioli, followed by the number of lungs from which they were taken, presented as bronchioli/lungs. (C) Recording of [Ca 2+ ] i in HBSMC with Fura-2 AM. Both β-NAD and ATP cause rise in [Ca 2+ ] i . n refers to the number of cells, taken from 4 independent experiments.

Article Snippet: HBSMC (C-12561, PromoCell, Heidelberg, Germany) were cultured in Smooth Muscle Cell Growth Medium 2 for at least 24 h at 37°C.

Techniques: Concentration Assay

Journal: PLOS One

Article Title: β-Nicotinamide adenine dinucleotide (β-NAD) acts as a bronchodilator

doi: 10.1371/journal.pone.0334491

Figure Lengend Snippet: (A, B) Recording of intracellular cAMP concentration in HBSMC via FRET, with low FRET ratio indicating high cAMP concentration. β-NAD and isoproterenol cause a decrease in FRET ratio, reflecting rise in intracellular cAMP concentration. In the presence of KH7 (30 µM), a soluble adenylyl cyclase antagonist, the cAMP response to β-NAD was blocked, while the isoproterenol-induced cAMP increase remained unaffected. (A) FRET ratio over time and (B) ΔFRET ratio (%) represents the change in response to β-NAD and isoproterenol, measured in the presence and absence of KH-7. Within-group comparisons include the ΔFRET ratio before agonist addition versus after the addition of β-NAD or isoproterenol. n represents the number of cells analyzed, derived from three independent experiments. Error bars indicate mean ± SEM throughout. Statistical analysis was performed using the Wilcoxon signed-rank test (two-tailed) with Bonferroni correction for multiple comparisons. (C-J) Force recording from trachea in organ bath (C-F) and videomorphometric recording of luminal bronchial area in PCLS (G-J) in specimens taken from mice lacking the C1 (C, E, G, I) or C2 domain of soluble adenylyl cyclase (D, F, H, J). In both assays, β-NAD-induced relaxation of muscarine-precontracted airways was not significantly reduced in knockout mice compared to their respective wildtype controls. (G-J) In PCLS, KH7 (30 µM) has no significant effect upon muscarine-induced contraction and β-NAD-induced relaxation, both in knockout and in wild-type mice. (E, F, I, J) Scatterplots depict changes induced by β-NAD related to the preceding response to muscarine. (E, F) Scatterplots show the β-NAD-induced relaxation effect (%) relative to the muscarine response. (I, J) Scatterplot showing the maximum peak responses of the second stimulation (first response set as 100%) in the presence of KH7 within C1 and C2 knockout groups and their corresponding wild-type controls. The corresponding controls with the application of vehicle (DMSO) instead of KH7 are depicted in . Statistical analysis was performed using the Mann-Whitney test. Data are expressed as mean ± SEM. n refers to the number of animals.

Article Snippet: HBSMC (C-12561, PromoCell, Heidelberg, Germany) were cultured in Smooth Muscle Cell Growth Medium 2 for at least 24 h at 37°C.

Techniques: Concentration Assay, Derivative Assay, Two Tailed Test, Knock-Out, MANN-WHITNEY

Journal: Nature Communications

Article Title: SARS-CoV-2 infection of human pluripotent stem cell-derived vascular cells reveals smooth muscle cells as key mediators of vascular pathology during infection

doi: 10.1038/s41467-024-54917-4

Figure Lengend Snippet: A Schematic of directed differentiation of hPSCs to vascular endothelial cells (ECs), smooth muscle cells (SMCs), and pericytes (PCs) (Created using BioRender). B Bulk RNA sequencing was performed on hPSC-derived ECs, PCs, and SMCs. The normalized expression values of known EC, PC, or SMC marker genes were quantitated over three biological replicates. The values plotted represent scaled normalized expression values for each gene across samples ( C ) (left) Principal component analysis (PCA) on bulk-RNA sequencing data from hPSCs, hPSC-ECs, hPSC-PCs, and hPSC-SMCs and primary endothelial cells (HUVECs), primary pericytes, and primary bronchial smooth muscle cells (BSMCs). (right) Mural cell-only PCA analysis (Primary pericytes, BSMCs, hPSC-PCs, and hPSC-SMCs) ( D ) Expression of PECAM1 and VE-Cadherin was quantitated by flow cytometry on hPSCs or on two independent differentiations of ECs. Data points on the bar graph represent values from two independent differentiations. Expression of VE-Cadherin, VWF, and PECAM1 in hPSC-derived ECs was observed by immunofluorescence. E Expression of NG2 was quantitated by flow cytometry on hPSCs or on two independent differentiations of PCs. Data points on bar graph represent values from two independent differentiations. Expression of PDGFR-B, and PDGFR-A was quantitated by flow cytometry on HEK293 cells, human dermal fibroblast (hDF) and two independent PC differentiation. Expression of SMA, PDGFR-B, and NG2 in hPSC derived PCs was observed by immunofluorescence. F Expression of PDGFR-B or SMA was quantitated by flow cytometry on hPSCs or on two independent differentiations of SMCs. Data points on the bar graph represent values from two independent differentiations. Expression of SMA, PDGFR-B, and NG2 in hPSC-derived SMCs was observed by immunofluorescence. Scale bar = 50 µm for all immunofluorescence images.

Article Snippet: Primary bronchial smooth muscle cells (BSMCs) were obtained from PromoCell (Cat# C12561).

Techniques: RNA Sequencing, Derivative Assay, Expressing, Marker, Flow Cytometry, Immunofluorescence