human vascular smooth muscle cells  (Cell Applications Inc)

Journal: bioRxiv

Article Title: Cardiovascular risk gene HDAC9 drives maladaptive vascular remodeling after arterial injury

doi: 10.64898/2026.05.12.723753

Figure Lengend Snippet: Ai-Aiv , Wire injury model. Hdac9 –/– Apoe –/– and Hdac9 +/+ Apoe –/– control littermate mice received Western-type diet one week before and for another three weeks after injury. Ai , Neointima formation was assessed in movat pentachrome-stained sections three weeks after injury. Scale bar represents 100 µm. Aii , Quantification of neointima area. n = 6-9 mice per genotype. Two-sided unpaired t test. Aiii , Representative DAPI and α-Sma staining. Aiv , Quantification of total cells in the neointima, and VSMC content. n = 3-4 mice per genotype. The quantification of cells is presented as the percentage of positively stained cells per section. Two-sided unpaired t test and Two-sided Mann-Whitney test. Scale bar represents 100 µm. Bi , Two-photon microscopy of endothelial Vcam-1 expression in ex vivo mounted carotid arteries. Hdac9 –/– Apoe –/– and Hdac9 +/+ Apoe –/– control littermate mice received Western-type diet for 4 weeks followed by imaging of Vcam-1 and Cd31 (n = 6-7 mice per group). Bi , Quantification of endothelial Vcam-1 in mounted carotid arteries. Scale bar, 50 µm. The quantification of Vcam-1 expression is presented as the percentage of endothelial lining per field of view. C , Quantitative proteomic profiling of MAoEC. Shown is the reactome pathway of differentially regulated proteins in TNF-α-stimulated cells (FDR-adj. p value < 0.05). Di, Dii , Transcriptomic profiling of mouse aortic smooth muscle cells (MAoSMC). MAoSMC, isolated from Hdac9 –/– Apoe –/– and Hdac9 +/+ Apoe –/– control mice, were stimulated with TNF-α for 6 hours and subjected to gene expression profiling. Di , Volcano plot of log 2 FoldChange ( Hdac9 +/+ Apoe –/– versus Hdac9 –/– Apoe –/– ) and –log 10 p-adjusted values showing top 30 differentially expressed genes related to cell migration. Highlighted in green and orange are the upregulated and downregulated genes respectively. Dii , Gene set enrichment analysis showing functional annotation of dysregulated genes (all FDR corrected). Diii , Scratch wound-healing assay. VSMC, isolated from Hdac9 –/– Apoe –/– and Hdac9 +/+ Apoe –/– control mice, were stimulated with PDGF-BB for 24 hours and analyzed for their migratory capacity. Quantification of the area covered by migrated cells. n = 5 mice per genotype. Two-sided unpaired t test. Div , Contractile markers and modulators of VSMC phenotypic transformation from the transcriptome analysis. Ei-Fiii , Functional and molecular assays in human aortic smooth muscle cells (HAoSMC). HAoSMC were transiently transfected with HDAC9 siRNA or scrambled control ( SCR ) RNA for 72 hours and subsequently stimulated with TNF-α (20 ng/mL) for indicated periods or left untreated. Ei , Quantification of PDGF-BB-induced proliferation of HAoSMCs by EdU assay. Eii , Representative immunoblots of VCAM-1 and Actin. Quantification of VCAM-1 normalized to Actin levels ( Eiii) , and IL-8 release ( Eiv). Fi , Representative immunoblots of p65 phosphorylation at serine 468 and total p65 levels. Quantification of phosphorylated p65 at serine 468 ( Fii ) and serine 536 ( Fiii ) normalized to total p65. n = 6-7 independent experiments. Two-way ANOVA with Bonferroni multiple comparison test for comparison of HDAC9 siRNA vs. SCR RNA. Data are means ± SEM.

Article Snippet: Human Aortic Smooth Muscle Cells (HAoSMCs) were purchased from PromoCell (Heidelberg, Germany), plated on cell culture dishes coated with collagen (Biochrom AG, Berlin, Germany) and cultured in smooth muscle cell growth medium (PromoCell, Heidelberg, Germany), according to manufacturer’s recommendations.

Techniques: Control, Western Blot, Staining, MANN-WHITNEY, Microscopy, Expressing, Ex Vivo, Imaging, Isolation, Gene Expression, Migration, Functional Assay, Wound Healing Assay, Transformation Assay, Transfection, EdU Assay, Phospho-proteomics, Comparison

Journal: bioRxiv

Article Title: Human platelet lysate drives clinically compliant generation of vascular mural cells from human pluripotent stem cells

doi: 10.64898/2026.02.03.703640

Figure Lengend Snippet: a) Representative immunofluorescent analysis of tube formation assay. Human umbilical venous endothelial cells (HUVEC) were co-cultured (5:1 ratio) with either day 13 human pluripotent stem cell (hiPSC) derived mural cells (MC) differentiated under human platelet lysate 1% (MC_hPL) or fetal bovine serum 10% (MC_FBS) supplemented conditions; or with human aortic smooth muscle cells (HAoSMC) as a positive control. HUVEC only cultures served as negative control. The left panel portrays the overall network formation (scale bar:100μm), while the right panel represents a magnified view of the same conditions (scale bar : 200μm) (n=3) . DAPI: 4′,6-diamidino-2-phenylindole. CNN1: Calponin 1. b)-e) Quantitative analysis of tube formation assay using Angiogenesis Analyzer in image J. Statistical analyses were performed using one-way ANOVA with Tukey’s post-hoc test and multiple comparison analysis (alpha value = 0,05) (n≥3). Significant differences (p<0,05) are indicated by bold parenthesis. b) Total Length (Tot. length) : Sum of all the branches and segments in the network. c) Total branching length (Tot. Branching length) cumulative length of all branches. d) Number of meshes (Nb meshes): indicator of network complexity by counting closed loops in the network. e) Number of Branches (Nb branches): indicator of sprouting potential.

Article Snippet: Tube formation assay was performed by seeding a cell suspension (2 × 10 5 cells/ml) composed of human venous umbilical endothelial cells (HUVEC) (PromoCell, Cat. No C-12200, Heidelberg, Germany) along with either human aortic smooth muscle cells (HAoSMC) (PromoCell, Cat. No C-12533), d13 derived mural cells either cultured with FBS or hPL1% supplementation, were seeded in a 5:1 ratio respectively on a Matrigel pre-coated μ-Slide 15 Well 3D Glass Bottom wells (Ibidi μ-Slide Angiogenesis, Cat. No. 81507.

Techniques: Tube Formation Assay, Cell Culture, Derivative Assay, Positive Control, Negative Control, Comparison