Journal: Toxicology Reports

Article Title: Effect of trivalent arsenicals on cell proliferation in mouse and human microvascular endothelial cells

doi: 10.1016/j.toxrep.2015.05.009

Figure Lengend Snippet: LC 50 values of trivalent arsenicals on microvascular endothelial cells.

Article Snippet: The mouse mammary fat pad microvascular endothelial cell line (MFP MVEC) was created from cells isolated from the mammary fat pad (MFP) tissue of two female H-2Kb-ts-A58 mice (Immortomice, Charles River Laboratories, Wilmington, MA) .

Techniques:

Journal: Cancers

Article Title: CAFs and TGF-β Signaling Activation by Mast Cells Contribute to Resistance to Gemcitabine/Nabpaclitaxel in Pancreatic Cancer

doi: 10.3390/cancers11030330

Figure Lengend Snippet: CM-HMC-1 and CM-CAF did not alter GEM/NAB-dependent inhibition of tumor angiogenesis but reduced the efficacy on tumor invasion. ( a ) Capillary morphogenesis. Both CM-HMC-1 and CM-CAF did not alter angiogenesis as demonstrated by microvascular formation and did not reduce the antiangiogenic potential of GEM/NAB. Pictures represent three different experiments. ( b ) Invasion assay. Both CM-HMC-1 and CM-CAF altered tumor invasion, by inducing the decrese and the increase of MIA PaCa-2 invasion, respectively. Both conditioned media reduced the efficacy of GEM/NAB-dependent inhibion of tumor invasion as reported in the histogram plot showing the recovery of tumor invasion on CM-HMC-1/CM-CAF + GEM/NAB treated cells. The pictures represent three different experiments.

Article Snippet: HMC-1 human mast cell line-1 cells were kindly provided by Prof. L. Macchia, University of Bari, CAF cells were purchased from Vitro Biopharma, and MVEC human dermal microvascular endothelial cells were purchased from Lonza, Switzerland.

Techniques: Inhibition, Invasion Assay

Journal:

Article Title: The neurotrophin receptor p75 NTR triggers endothelial cell apoptosis and inhibits angiogenesis: implications for diabetes-induced impairment of reparative neovascularization

doi: 10.1161/CIRCRESAHA.108.177386

Figure Lengend Snippet: (a) HUVEC were infected with 100, 250 and 500 M.O.I. of Ad.Null or Ad.p75NTR, as indicated. After 48h, cell lysates were collected and subjected to western blotting with antibodies to p75NTR, cleaved caspase-3, and GAPDH (used as loading control). (b) FACS analyses for p75NTR show abundant receptor expression (95.8±3%) at 12h from gene transfer with Ad.p75NTR (250 M.O.I.), while at 12h from Null gene transfer, p75NTR-expressing HUVEC are only 6.7±0.7%. (c) p75NTR-expressing HUVEC were gated and studied at 12h, 24h, and 48h for co-expression with Annexin-V and propidium iodide (PI) to detect apoptosis. This analysis revealed that at 12h from Ad.p75NTR, less than 9% of p75NTR-carrying cells presented Annexin-V on the external plasma membrane. Early apoptosis (cells positive for Annexin-V and negative for PI, lower right squares) peaked at 24h (39.1±3% of p75NTR-expressing HUVEC), followed by late apoptosis (cells positive for both Annexin-V and PI, upper right squares) at 48h from Ad.p75NTR (17.05±2% of p75NTR-expressing HUVEC). (d) HUVEC were treated as described in (a) and apoptotic nuclei were detected by TUNEL assay. Fluorescent images are representative of apoptosis rate in Null-HUVEC and p75NTR-HUVEC. Bar graphs quantify apoptosis, which is expressed as percentage of TUNEL-positive nuclei (green fluorescence) to total nuclei (stained in blue fluorescence by DAPI). Data are presented as means±SEM. *P<0.05 and **P<0.001 vs. Ad.Null. (e,f) Caspase-3 activity assay was performed on HUVEC (e) or MVEC (f). Cells (5000 cells/well) were infected with 100, 250 or 500 M.O.I. of Ad.Null or Ad.p75NTR or left uninfected (PBS). After 48h, Caspase-Glo 3/7 was incubated for 1h before recording luminescence. The apoptosis inducer staurosporin (stauro, 1μM) was used as reference in (e). Values are means±SEM. *P<0.05 and **P<0.001 vs. Ad.Null. (g) Upper panel: Concentration of endothelial apoptotic microparticles (EMP) released by Null-HUVEC or p75NTR-HUVEC under no other stimulation or following incubation with the apoptosis inducer staurosporin (100 nM, 16h). Values are means±SEM. **P<0.01 vs. Ad.Null. Lower panel: p75NTR is present in EMP released by HUVEC. The shadow peak corresponds to fluorescence background obtained with the isotypic control of p75NTR antibody. The second peak represents the specific labelling of EMPs with fluorescent p75NTR antibody. (h) Caspase-3 activity assay performed on HUVEC infected with Ad.Null or Ad. p75NTR (each at 100 and 250 M.O.I.) before being treated with PBS, proNGF (5ng/mL), NGF (100ng/mL), or BDNF (100ng/mL) for 24h. Values are means±SEM. *P<0.01 vs. PBS.

Article Snippet: HUVEC and MVEC were purchased from Cambrex (Belgium) and grown in EGM-2 medium (EBM-2 supplemented with EGM-2 SingleQuots, Cambrex) containing 5% FBS (Cambrex).

Techniques: Infection, Western Blot, Control, Expressing, Clinical Proteomics, Membrane, TUNEL Assay, Fluorescence, Staining, Caspase-3 Activity Assay, Incubation, Concentration Assay

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Endothelial sprouting in bulk fibrin hydrogels. A) Fluorescent images of embedded MVEC stained red with UEA-1 in gels made with different MVEC:FB ratios and cultured in different medium volumes. B) Quantitation of the average sprout length in bulk gels (n=3). C) Quantitation of the number of sprouts >100 µm in length in bulk gels (n=3). Error bars represent standard deviation of the mean. * indicates statistical significance (*p<0.05, **p<0.01 and ***p<0.001).

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques: Staining, Cell Culture, Quantitation Assay, Standard Deviation

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Influence of cell ratios and culture media volume on total sprouts.

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques:

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Influence of cell ratios and culture media volume on average sprout length.

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques:

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Characterization of modular microtissues. A) Single MVEC:FB microtissues (1:1 and 1:3) under phase contrast showing microtissue morphology and embedded cells immediately after microbead fabrication. B) Single MVEC:FB microtissues (1:1 and 1:3) under fluorescence showing cell viability (green = live cells, red = dead cells). C) Quantitation of cell viability for a population of microtissues. D) Quantitation of total cells per unit volume for a population of microtissues. E) Total DNA in microtissues as a function of time. Best viewed in color. Error bars represent standard deviation of the mean. * indicates statistical significance (*p<0.05).

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques: Fluorescence, Quantitation Assay, Standard Deviation

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Flow cytometry analysis of MVEC and FB cell population in fibrin co-cultures over time. A,B) Mono-culture of MVEC and FB. C- H) MVEC and FB cell population in co-cultures over time. Error bars represent standard deviation of the mean. * indicates statistical significance (*p<0.01).

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques: Flow Cytometry, Standard Deviation

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Sprouting of neovessels from modular microtissues. A) Fluorescence images (green = fibrin, red = MVEC) at day 7 and 14 with different MVEC:FB ratios. B) Quantification of vessel area normalized to microtissue area. C) Box plot of vessel diameter. The solid center line in the box plot represents the median, the dotted center line in the box represents the mean, and the lower and upper boundaries of the box represent the 25th and 75th percentiles, respectively. Whiskers (error bars) above and below the box indicate the 90th and 10th percentiles. Large dots represent outliers.

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques: Fluorescence

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Vessel lumen diameter at day 7 and day 14 in embedded microtissue cultures

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques:

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Inosculation of MVEC neovessels. A) MVEC (red) networks (i) sprouted from microtissues (green) and formed branches (ii) with hollow lumens (iii, iv). B) Serial histological sections showed inosculation of adjacent MVEC vessels.

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques:

Journal: ACS biomaterials science & engineering

Article Title: Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

doi: 10.1021/acsbiomaterials.6b00274

Figure Lengend Snippet: Microtissues cultured in suspension aggregated to form larger tissue structures. A, B) By day 7, neovessels were evident in tissue masses. C, D) By day 14, networks of vessels had formed in tissue masses made with 1:3 MVEC:FB microtissues, but were less evident in those made at 1:1. E) Fluorescence staining and F) PECAM/CD-31 IHC confirmed that vessels within tissue structures were created by MVEC.

Article Snippet: Human neonatal foreskin microvascular endothelial cells (MVEC) and normal human lung fibroblasts (FB, passage 9–12) were obtained from a commercial vendor (Lonza Inc., Walkersville, MD).

Techniques: Cell Culture, Suspension, Fluorescence, Staining