Journal: Tissue Engineering. Part A

Article Title: Endothelial Cell Response to Chemical, Biological, and Physical Cues in Bioactive Hydrogels

doi: 10.1089/ten.tea.2013.0602

Figure Lengend Snippet: Effects of protein concentration, functionalization density, and type on BAOEC adhesion over 1 week. ANOVA analysis revealed significant effects of all protein variables (concentration, functionalization density, and type) and time on adhesion levels (p<0.001). Tukey's post hoc test was applied for independent variables with greater than two levels with significance noted by the following: *p<0.001 relative to all other time points for given protein type; **p<0.003 relative to the 3-h time point for given protein type; †p<0.001 relative to all other concentrations of given protein type; •p<0.001 relative to all other functionalization densities of given protein type. n=4 samples, 3 images per sample for a total of 12 images; measurements are expressed as mean±standard error. BAOECs, bovine aortic endothelial cells. Color images available online at www.liebertpub.com/tea

Article Snippet: Initial protein incorporation (I) was measured from PR as follows: EC adhesion and spreading Bovine aortic endothelial cells (BAOECs; Cell Applications, Inc.) were used for all cell studies.

Techniques: Protein Concentration, Concentration Assay

Journal: Tissue Engineering. Part A

Article Title: Endothelial Cell Response to Chemical, Biological, and Physical Cues in Bioactive Hydrogels

doi: 10.1089/ten.tea.2013.0602

Figure Lengend Snippet: Effects of protein concentration and functionalization density on BAOEC (A) migration and (B) 3-h spreading on PEG-Scl2-2 and PEG-collagen gels. (C) Representative images of BAOECs adhered to samples at 3 h. Scale bar applied to all images. (A) n=3 samples, 9 images per sample for a total of 27 images; (B) n=3 samples, 3 images per sample for a total of 9 images; measurements are expressed as mean±standard error; *p<0.1 relative to all other samples; †p<0.05 relative to Scl2-2 1×, 8 mg/mL sample; ‡p<0.05 relative to collagen and Scl2-2 0.1×, 1 mg/mL samples. PEG, poly(ethylene glycol). Scl2-2, Streptococcal collagen like. Color images available online at www.liebertpub.com/tea

Article Snippet: Initial protein incorporation (I) was measured from PR as follows: EC adhesion and spreading Bovine aortic endothelial cells (BAOECs; Cell Applications, Inc.) were used for all cell studies.

Techniques: Protein Concentration, Migration

Journal: Pharmaceutical Research

Article Title: Pharmacokinetics and Pharmacodynamic Effect of a Blood-Brain Barrier-Crossing Fusion Protein Therapeutic for Alzheimer’s Disease in Rat and Dog

doi: 10.1007/s11095-022-03285-z

Figure Lengend Snippet: FC5 binding to canine, rat and human endothelial cells. Binding studies were carried out as described in Methods section. Bar graph represents data from two independent studies and expressed as mean and range. CnAOEC: Canine (dog) aortic endothelial cells; SvRBEC: rat brain endothelial cells; hCMEC/D3: human brain endothelial cells. MFI: Mean Fluorescence Intensity. Since the objective of this experiment was simply to demonstrate cross-reactivity of FC5 with canine endothelial cells as a prerequisite for PK studies, binding across the three cell types was not compared statistically.

Article Snippet: Immortalized human (HCMEC/D3/HBEC-D3, Dr. Pierre Olivier Couraud, Cochin Institute Université Paris Descartes INSERM, Ref ), rat microvascular brain endothelial cells (sv-ARBEC) and Canine Aortic Endothelial Cells (CnAOEC, Cell Applications Inc., San Diego, CA, USA) were cultured until confluent at 37°C in a humidified 5% CO2 atmosphere in T-75 flasks coated with 100 μg/ml rat tail collagen type 1 (BD Canada, Mississauga, ON, Canada).

Techniques: Binding Assay, Fluorescence

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Pleiotrophin Induces Transdifferentiation of Monocytes Into Functional Endothelial Cells

doi: 10.1161/01.ATV.0000222017.05085.8e

Figure Lengend Snippet: PTN downregulates expression of monocytic cell markers. A, Total RNA was extracted from THP-1 cells grown in 10% serum (lane 2), induced to differentiate into macrophage-like cells by addition of 25 ng/mL phorbol 12-myristate 13-acetate (PMA; lane 3), transduced with retroviral bicistronic vector expressing: GFP (lane 4), PTN sense strand (lane 5), PTN antisense strand (lane 6) followed by treatment with PMA. The exponentially growing human coronary artery endothelial cells (lane 7) were used as a negative control. Analyzed monocytic cell markers were c-fms and CD-68 with primers predicted to amplify 97- and 132-bp DNA fragments, respectively. GAPDH primers were used as control a for the RT-PCR. Lane 1 is a DNA ladder marker. B, Flow cytometry analysis was performed by incubating 5×105 THP-1 cells expressing PTN or GFP with phycoerythrin-labeled anti-CD14 antibody from PharMingen. Human coronary artery endothelial cells were used as a negative control. Uninfected THP-1 cells were used as positive control, and human coronary endothelial cells (endothelial) were used as a negative control. FACS analysis was performed at the Cedars-Sinai Research Institute Core Facility. Each experiment was repeated 3 times, and each bar graph represents mean±SEM of 3 experiments.

Article Snippet: Human aortic endothelial cells and smooth muscle cells were obtained from Cell Applications, INC. and cultured according to their recommendation.

Techniques: Expressing, Transduction, Retroviral, Plasmid Preparation, Negative Control, Control, Reverse Transcription Polymerase Chain Reaction, Marker, Flow Cytometry, Labeling, Positive Control

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Pleiotrophin Induces Transdifferentiation of Monocytes Into Functional Endothelial Cells

doi: 10.1161/01.ATV.0000222017.05085.8e

Figure Lengend Snippet: PTN upregulates expression of endothelial cell markers in the monocytic cells. A, RT-PCR analysis of endothelial cell markers. Total RNA isolated from untransduced and transduced cells were used for RT-PCR analysis, using specific primers for endothelial cell markers (see online supplement). To ensure semiquantitative results of the RT-PCR analysis, the number of PCR cycles for each set of primers was checked to be in the linear range of the amplification. In addition, all RNA samples were adjusted to yield equal amplification of GAPDH as an internal standard. The amplified products were separated on 1.2% agarose gels and stained with ethidium bromide. B, A representative flow cytometry analysis of αvβ3 integrin expression by the monocytic cells. GFP (top left panel) or PTN-expressing THP-1 cells (bottom left panel) were incubated with a 1:100 dilution of anti-human αvβ3 mouse antibody (Chemicon Co). After washing, cells were incubated with 1:500 dilution of phycoerythrin-labeled anti-mouse antibody (Sigma), fixed, and analyzed by FACS, as described above. In addition, human coronary artery endothelial cells in the absence (top right panel) or presence of anti-αvβ3 antibody (bottom right panel) were used as a positive control. C, RT-PCR analysis of transcription factors. The PCR was performed as described in A, and the primers and PCR condition are described in the online supplement.

Article Snippet: Human aortic endothelial cells and smooth muscle cells were obtained from Cell Applications, INC. and cultured according to their recommendation.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Amplification, Staining, Flow Cytometry, Incubation, Labeling, Positive Control

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Pleiotrophin Induces Transdifferentiation of Monocytes Into Functional Endothelial Cells

doi: 10.1161/01.ATV.0000222017.05085.8e

Figure Lengend Snippet: Amplification plot for the expression of CD68 expression in the monocytic cells. Oligonucleotide primer pairs for CD68 gene and an oligonucleotide probe labeled with a reporter fluorescent dye at the 5′ end and quencher dye at the 3′ end were designed using Oligo 4.0 software (National Bioscience, Plymouth, MN). Total RNA with DNase I treatment was used to synthesize first-stand cDNA with RT (GIBCOBRL) and oligo(dT) 15 Primer (Promega). Total RNA (50 ng) was added to a 50 μl reverse transcriptase-polymerase chain reaction (RT-PCR) reaction mixture according to the manufacturer’s protocol (Roche Molecular Systems). The products of the RT reactions was used to seed real-time PCR by using an ABI Prism 7700 Sequence Detector by comparing with GAPDH (internal control) and individual standard curve performed in triplicate. The thermal cycling conditions included one cycle at 48°C for 30 minutes, one cycle at 95°C for 10 minutes, 40 cycles at 95°C for 15 s, annealing at 60°C for 1 minute, and a final hold at 25°C for 2 minutes. Standard curves for the expression of each gene were generated by serial dilution of a standard preparation of total RNA isolated from cultured monocytic or endothelial cells.

Article Snippet: Human aortic endothelial cells and smooth muscle cells were obtained from Cell Applications, INC. and cultured according to their recommendation.

Techniques: Amplification, Expressing, Labeling, Software, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Sequencing, Control, Generated, Serial Dilution, Isolation, Cell Culture

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Pleiotrophin Induces Transdifferentiation of Monocytes Into Functional Endothelial Cells

doi: 10.1161/01.ATV.0000222017.05085.8e

Figure Lengend Snippet: Quail chorioallantotic membrane (CAM) assay. Fertilized eggs of Japanese quail were cultured at 37 °C under ambient atmosphere, for 56 hours, and then opened at embryonic day 3 (panel A) after incubation of the eggs and cultured further at 37°C in 6-well plates. At E7 (Panel B), 1×106 RAW cells in prewarmed PBS (200 μl) were applied in drops to the surface of each CAM. The embryos were incubated further at 37°C for 72 hours, at which time they were harvested (panel C), fixed in 4% paraformaldehyde/2% glutaraldehyde/PBS. Panel D and E show fluorescent Image of CAM implanted with RAW cells expressing PTN/GFP or mouse endothelial cells expressing GFP, respectively.

Article Snippet: Human aortic endothelial cells and smooth muscle cells were obtained from Cell Applications, INC. and cultured according to their recommendation.

Techniques: Membrane, Chick Chorioallantoic Membrane Assay, Cell Culture, Incubation, Expressing

Journal: Molecules

Article Title: Expanding the Reactive Sulfur Metabolome: Intracellular and Efflux Measurements of Small Oxoacids of Sulfur (SOS) and H 2 S in Human Primary Vascular Cell Culture

doi: 10.3390/molecules26237160

Figure Lengend Snippet: Intracellular endogenous concentrations of analytes in ( A ) human aortic/coronary smooth muscle cells and ( B ) human aortic/coronary endothelial cells. * = p < 0.05; ** = p < 0.01; *** = p < 0.001; **** = p < 0.0001; ns = not significant.

Article Snippet: Human aortic endothelial cells were purchased from iXCells Biotech (San Diego, CA, USA) (10HU-020, Mixed Donors), and human aortic smooth muscle cells were purchased from Gelantis (San Diego, CA, USA) (PH35405A, Donor Age: 33, Donor Sex: Male).

Techniques:

Journal: Molecules

Article Title: Expanding the Reactive Sulfur Metabolome: Intracellular and Efflux Measurements of Small Oxoacids of Sulfur (SOS) and H 2 S in Human Primary Vascular Cell Culture

doi: 10.3390/molecules26237160

Figure Lengend Snippet: Rates of efflux of analytes averaged over three 1 h periods in ( A ) human aortic/coronary smooth muscle cells and ( B ) human aortic/coronary endothelial cells. * = p < 0.05; ** = p < 0.01; **** = p < 0.0001; ns = not significant.

Article Snippet: Human aortic endothelial cells were purchased from iXCells Biotech (San Diego, CA, USA) (10HU-020, Mixed Donors), and human aortic smooth muscle cells were purchased from Gelantis (San Diego, CA, USA) (PH35405A, Donor Age: 33, Donor Sex: Male).

Techniques:

Journal: Molecules

Article Title: Expanding the Reactive Sulfur Metabolome: Intracellular and Efflux Measurements of Small Oxoacids of Sulfur (SOS) and H 2 S in Human Primary Vascular Cell Culture

doi: 10.3390/molecules26237160

Figure Lengend Snippet: ( A ) Oxygen consumption rate ( J O 2 ). ( B ) H 2 O 2 production rate in human aortic smooth muscle cells during normoxic (N) and hypoxic (H) growth. ( C ) Western blot showing protein expression of cellular SQOR with GAPDH used as loading control. ( D ) Fold change of SQOR protein expression for normoxia versus hypoxia. ** = p < 0.01; *** = p < 0.001; ns = not significant.

Article Snippet: Human aortic endothelial cells were purchased from iXCells Biotech (San Diego, CA, USA) (10HU-020, Mixed Donors), and human aortic smooth muscle cells were purchased from Gelantis (San Diego, CA, USA) (PH35405A, Donor Age: 33, Donor Sex: Male).

Techniques: Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: An Engineered Gene Nanovehicle Developed for Smart Gene Therapy to Selectively Inhibit Smooth Muscle Cells: An In Vitro Study

doi: 10.3390/ijms21041530

Figure Lengend Snippet: hEGR1 promoter activities in ( A ) smooth muscle cells (SMCs) and ( B ) endothelial cells (ECs) were characterized using the Lucia gene as a report gene. phEGR1-Lucia was transfected by Effectene ® Transfection Reagent, a commercial gene vehicle, to eliminate the noise factors from vehicles.

Article Snippet: Rat aortic endothelial cells were purched from Angio-Proteomie, USA.

Techniques: Transfection

Journal: International Journal of Molecular Sciences

Article Title: An Engineered Gene Nanovehicle Developed for Smart Gene Therapy to Selectively Inhibit Smooth Muscle Cells: An In Vitro Study

doi: 10.3390/ijms21041530

Figure Lengend Snippet: Western blot was used to evaluate the amount of functional protein (protein kinase C-delta, PKCδ) expressed in ( A ) smooth muscle cells and ( B ) endothelial cells transfected with the phEGR1-PKCδ gene using Effectene ® Transfection Reagent as the gene vehicle. Control groups are wild type cells which were not treated with phEGR1-Lucia or phEGR-PKCδ genes.

Article Snippet: Rat aortic endothelial cells were purched from Angio-Proteomie, USA.

Techniques: Western Blot, Functional Assay, Transfection, Control

Journal: International Journal of Molecular Sciences

Article Title: An Engineered Gene Nanovehicle Developed for Smart Gene Therapy to Selectively Inhibit Smooth Muscle Cells: An In Vitro Study

doi: 10.3390/ijms21041530

Figure Lengend Snippet: The in vitro inhibition effect of the PEI-Au/CHC/phEGR1-PKCδ complex on the smooth muscle cells and endothelial cells was evaluated with PrestoBlue ® reagent at ( A ) 24 h and ( B ) 48 h after transfection. The cell viability of endothelial cells and smooth muscle cells treated with differing PEI-Au/CHC/DNA complexes was assessed. Cells were treated with lipopolysaccharides (LPS) to simulate the inflammatory environment. ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Rat aortic endothelial cells were purched from Angio-Proteomie, USA.

Techniques: In Vitro, Inhibition, Transfection