pcs 100 013  (ATCC)

Journal: Biology

Article Title: Endothelial PAI-1 Drives Lead-Induced Cerebral Amyloid Angiopathy via Activation of C3 + Decorin + A1-like Astrocytes

doi: 10.3390/biology15040297

Figure Lengend Snippet: Pb-treated endothelial cells promote the activation of C3 + decorin + SVG astrocytes. ( A ) Representative SVG astrocytes immunostained for GFAP (green), C3 (red), and decorin (magenta) after incubation with HUVEC-CM from control or Pb-treated HUVEC. Arrowheads indicate positive C3 + decorin + A1-like astrocytes. ( B ) Quantification of triple-positive C3 + decorin + A1-like astrocytes. Data are demonstrated as mean ± SD, *: p < 0.05, analyzed by one-way ANOVA with Dunnett’s post hoc test. Note: Control: sodium acetate-treated group, Pb: lead acetate-treated group.

Article Snippet: This study used human umbilical vein endothelial cells (HUVECs; PCS-100-013, <20 passages) and human SVG p12 astrocytes (CRL-8621) [ ], as well as cells obtained from the mouse endothelial cell line bEnd.3 (CRL-2299) and the mouse astrocyte cell line C8-D1A (CRL-2541) (all purchased from ATCC, Manassas, VA, USA).

Techniques: Activation Assay, Incubation, Control

Journal: Biology

Article Title: Endothelial PAI-1 Drives Lead-Induced Cerebral Amyloid Angiopathy via Activation of C3 + Decorin + A1-like Astrocytes

doi: 10.3390/biology15040297

Figure Lengend Snippet: Pb exposure upregulates endothelial PAI-1 expression in vitro. ( A ) Representative Western blot showing PAI-1 protein expression in HUVECs treated with control (NaAc) or 1 µM PbAc. Uncropped Western blot images shown in supplement. ( B ) Quantification of PAI-1 protein levels in HUVEC lysates normalized to GAPDH ( n = 6). ( C ) PAI-1 concentrations in HUVEC-conditioned media (HUVEC-CM) with or without immunodepleting of PAI-1 or IgG control measured by ELISA. Data are demonstrated as mean ± SD; *: p < 0.05, analyzed by one-way ANOVA with Dunnett’s post hoc test. Note: Ctrl: control (sodium acetate), Pb: lead acetate, CM: conditioned media, IP: immunodepleting of PAI-1, PAI-1: plasminogen activator inhibitor-1.

Article Snippet: This study used human umbilical vein endothelial cells (HUVECs; PCS-100-013, <20 passages) and human SVG p12 astrocytes (CRL-8621) [ ], as well as cells obtained from the mouse endothelial cell line bEnd.3 (CRL-2299) and the mouse astrocyte cell line C8-D1A (CRL-2541) (all purchased from ATCC, Manassas, VA, USA).

Techniques: Expressing, In Vitro, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Free-floating long-term vascularized mesenchymal organoids

doi: 10.1016/j.isci.2025.114548

Figure Lengend Snippet: VMOs formed at different EC:MSC ratios support long-term culture and preserve morphology (A) Schematic of the vascular mesenchymal organoid (VMO) formation process, combining human umbilical vein endothelial cells (HUVECs) and mesenchymal stem/stromal cells (MSCs) at 25:75, 50:50, and 75:25 ratios with Matrigel to generate organoids, no-Matrigel spheroid aggregates were cultured for comparison. (B) Time-lapse images show organoid formation during the first 3 days across different endothelial:MSC ratios. Organoids are fully formed by day 3 (D3) in all tested ratios. (C) Whole mount immunofluorescence analysis of D14 VMOs with different seeding ratios, demonstrating the presence of both VE-Cadherin + endothelial cells and CD90 + MSCs. (D) Circularity measurements of organoids over 60 days of culture, showing stable (>0.8) circularity across all ratios. (E) Quantification of the cross-sectional area of organoids at different seeding ratios, showing that the 50:50 ratio produces the largest organoid area. (F) Analysis of luminescent cell viability signal over a 60-day culture period, revealing that the 50:50 ratio exhibits the highest proliferative capacity during the first 35 days, while the 25:75 ratio had the fewest cells throughout the culture duration. Two-way ANOVA with Šidák’s multiple comparisons test was used, and asterisks denote statistical significance: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Scale bars 50 μm (C) and 100 μm (B). (D–F) show mean values ± SD with n = 16 individual VMO. See also .

Article Snippet: Human umbilical vein endothelial cells (HUVEC) , ATCC , PCS-100-013.

Techniques: Cell Culture, Comparison, Immunofluorescence

Journal: JACC: Basic to Translational Science

Article Title: Estrogen Influences Human Microvascular Endothelial Function Via Sex-Specific Regulation of Sphingolipids

doi: 10.1016/j.jacbts.2025.101389

Figure Lengend Snippet: Chronic Estrogen Exposure Results in Ceramide Accumulation and Promotes the Formation of Endothelial H 2 O 2 Formation H 2 O 2 fluorescence, as measured using peroxy-yellow-1 (PY1) in human umbilical vein endothelial cells treated with 1 and 100 nmol/L 17β-estradiol (E2) (48 hours) in (A) women (control n = 10, 1 nmol/L E2 n = 8, 100 nmol/L E2 n = 10) and (B) men (control n = 15, 1 nmol/L E2 n = 7, 100 nmol/L E2 n = 11); (C) Sex difference. H 2 O 2 in endothelial cells treated with 100 nmol/L E2 +/− catalase (500 U/ml) in (D) women (control n = 10, E2 n = 10, E2+catalase n = 7) and (E) men (control n = 15, E2 n = 11, E2+catalase n = 5). H 2 O 2 production in the presence of E2 vs E2+ GW4869 (4 μmol/L; 48 hours) in endothelial cells from (F) women (control n = 10, E2 n = 10, E2+GW4869 n = 9) and (G) men (control n = 15, E2 n = 11, E2+GW4869 n = 11). Endothelial H 2 O 2 following exposure to C2-ceramide (10 μmol/L, 48 hours) in (H) women (control n = 10, ceramide n = 6) and (I) men (control n = 15, ceramide n = 7). Data are presented as mean ± SEM. (A, B, D-G) 1-way analysis of variance with Holm-Sidak multiple comparisons test and (C, H-I) 2-tailed t -test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Human umbilical vein endothelial cells (ATCC Manassas) from 3 women and 3 men were pooled by biological sex and cultured using 5% fetal bovine serum containing endothelial cell growth medium (EBM-2 Basal Medium; Lonza CC-3156).

Techniques: Fluorescence, Control

Journal: JACC: Basic to Translational Science

Article Title: Estrogen Influences Human Microvascular Endothelial Function Via Sex-Specific Regulation of Sphingolipids

doi: 10.1016/j.jacbts.2025.101389

Figure Lengend Snippet: Endothelial Ceramide Content Is Higher Among Men Compared With Women at Baseline and Following Chronic Estrogen Exposure Normalized endothelial (A) total, (B) medium-chain, (C) long-chain, and (D) very-long chain ceramide levels in endothelial cells from men and women. (E) Total, (F) medium-chain, (G) long-chain, and (H) very-long chain ceramide levels in endothelial cells from women following vehicle (EtoH %vol/vol), 1 or 100 nmol/L 17β-estradiol (E2; 48 hours) treatment. (I) Total, (J) medium-chain, (K) long-chain, and (L) very-long chain ceramide levels in endothelial cells from men following vehicle, 1 and 100 nmol/L E2 treatment. Percent change from vehicle in (M) total, (N) medium-chain, (O) long-chain, and (P) very-long chain ceramide levels in endothelial cells from women compared with men following 1 and 100 nmol/L E2 treatment. Data are presented as mean ± SEM. (A-D) 2-tailed t -test, (E-P) 1-way analysis of variance ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Human umbilical vein endothelial cells (ATCC Manassas) from 3 women and 3 men were pooled by biological sex and cultured using 5% fetal bovine serum containing endothelial cell growth medium (EBM-2 Basal Medium; Lonza CC-3156).

Techniques:

Journal: JACC: Basic to Translational Science

Article Title: Estrogen Influences Human Microvascular Endothelial Function Via Sex-Specific Regulation of Sphingolipids

doi: 10.1016/j.jacbts.2025.101389

Figure Lengend Snippet: There Are Sex Differences in Ceramide to S1P Ratios at Baseline and Following Estrogen Treatment Normalized endothelial (A) sphingosine and (B) sphingosline-1-phosphate (S1P) levels as well (C) total ceramide to S1P ratio in control cells from men and women. (D) Sphingosine and (E) S1P levels as well (F) total ceramide to S1P ratio in endothelial cells from women following vehicle (EtoH %vol/vol), 1 or 100 nmol/L 17β-estradiol (E2) (48 hours) treatment. (G) Sphingosine and (H) S1P levels as well (I) total ceramide to S1P ratio in endothelial cells from women following vehicle, 1 or 100 nmol/L E2 treatment. Data are presented as mean ± SEM. (A-C) 2-tailed t -test, (D-I) 1-way analysis of variance ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Human umbilical vein endothelial cells (ATCC Manassas) from 3 women and 3 men were pooled by biological sex and cultured using 5% fetal bovine serum containing endothelial cell growth medium (EBM-2 Basal Medium; Lonza CC-3156).

Techniques: Control

Journal: JACC: Basic to Translational Science

Article Title: Estrogen Influences Human Microvascular Endothelial Function Via Sex-Specific Regulation of Sphingolipids

doi: 10.1016/j.jacbts.2025.101389

Figure Lengend Snippet: MitoTempol Protects Against Estrogen-Induced Endothelial Dysfunction (A) FID in vessels from men treated with 100 nmol/L 17β-estradiol (E2) (16-20 hours; n = 6) vs E2+MitoTempol (100 μmol/L; n = 4). (B) FID following E2+MitoTempol (100 μmol/L; n = 5) treatment within microvessels from women that had impaired dilation to flow following 100 nmol/L 17β-estradiol (E2) (16-20 hours; n = 5). Among vessels that maintained dilation to flow with 100 nmol/L E2 alone, dilation to flow following E2+MitoTempol (100 μmol/L; n = 3) treatment (C) +/− 30 minutes catalase (500 U/mL; n = 3), (D) +/− 30 minutes L-NAME (100 μmol/L; n = 3). H 2 O 2 in HUVECs from (E) women (control n = 10, E2 n = 10, E2+MitoTempol n = 6) and (F) men (control n = 15, E2 n = 11, E2+MitoTempol n = 8) treated with 100 nmol/L 17β-estradiol (E2; 48 hours) +/− MitoTempol (100 μmol/L). (G) Schematic of results. Data are presented as mean ± SEM. (A-E) 2-way analysis of variance and (F-G) 1-way analysis of variance Holm-Sidak multiple comparisons test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Human umbilical vein endothelial cells (ATCC Manassas) from 3 women and 3 men were pooled by biological sex and cultured using 5% fetal bovine serum containing endothelial cell growth medium (EBM-2 Basal Medium; Lonza CC-3156).

Techniques: Control