Journal: Pharmacological research

Article Title: Modulation of cellular bioenergetics by CO-releasing molecules and NO-donors inhibits the interaction of cancer cells with human lung microvascular endothelial cells.

doi: 10.1016/j.phrs.2018.09.005

Figure Lengend Snippet: Fig. 4. Effect of CORM-401 and PAPA NONOate on MDA-MB-231-luc2-tdTomato cell transendothelial migration and adhesion to lung microvascular endothelial cells.

Article Snippet: Human lung microvascular endothelial cell line (hLMVEC) was purchased from the European Cell Culture Collection (Cell Applications, San Diego, CA, USA), human breast adenocarcinoma MDA-MB-231-luc2-tdTomato cell line stably expressing the firefly luciferase gene and tdTomato fluorescent protein was kindly provided by Prof. Joanna Wietrzyk (Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences). hLMVEC cells were maintained in Microvascular Endothelial Cell Growth Medium (Cell Applications), MDA-MB-231-luc2-tdTomato cells were cultured in RPMI1640 GlutaMAX Medium (Gibco) with 10% FBS (Gibco), AAS (Sigma-Aldrich) containing 20 units of penicillin, 20 mg streptomycin and 0.05 mg amphotericin B.

Techniques: Migration

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: GSK-3Beta-Dependent Activation of GEF-H1/ROCK Signaling Promotes LPS-Induced Lung Vascular Endothelial Barrier Dysfunction and Acute Lung Injury

doi: 10.3389/fcimb.2017.00357

Figure Lengend Snippet: LPS induces GSK-3beta activation in dose- and time-dependent manners in HPMECs. Expression of P-GSK-3beta and GSK-3beta was detected after incubation with different concentrations of LPS for 1 h (A) . The expression of P-GSK-3beta was represented as a histogram according to band intensities (B) . Expression of P-GSK-3beta and GSK-3beta was examined at indicated time points after stimulation with LPS (0.1 μg/ml) in HPMECs (C) . The Western blotting results are presented as a histogram showing the band intensity values (D) . * P < 0.05 vs. LPS un-treatment group.

Article Snippet: Primary human pulmonary micro-vascular endothelial cells (HPMECs) were obtained ScienCell Research Laboratories and maintained in ScienCell Endothelial Cell Medium in a humidified 37°C, 5% CO 2 incubator.

Techniques: Activation Assay, Expressing, Incubation, Western Blot

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: GSK-3Beta-Dependent Activation of GEF-H1/ROCK Signaling Promotes LPS-Induced Lung Vascular Endothelial Barrier Dysfunction and Acute Lung Injury

doi: 10.3389/fcimb.2017.00357

Figure Lengend Snippet: Involvement of GSK-3beta in LPS-induced GEF-H1/ROCK signaling activation. HPMECs were incubated with LPS (0.1 μg/ml) at different indicated times, and the GEF-H1 and myosin-associated phosphatase type 1 (P-MYPT 1: the substrate of ROCK) were detected by Western blot assay (A) . The expression of GEF-H1 and P-MYPT 1 were represented as a histogram according to band intensities (B) . * < 0.05 vs. LPS un-treatment group. Inhibition effect of GSK-3beta activity in HPMECs was analyzed by Western blot (C,D) . * P < 0.05 vs. the negative control group, # P < 0.05 vs. the corresponding LPS treatment group. HPMECs were pretreated with SB-216763 (20 μM) for 1 h and then were exposed to LPS (0.1 μg/ml) for 1 h. The expression of GEF-H1 and P-MYPT 1 were determined by Western blot (E) . The Western blotting results are presented as a histogram showing the band intensity values (F) . * P < 0.05 vs. the negative control group, # P < 0.05 vs. the corresponding LPS treatment group.

Article Snippet: Primary human pulmonary micro-vascular endothelial cells (HPMECs) were obtained ScienCell Research Laboratories and maintained in ScienCell Endothelial Cell Medium in a humidified 37°C, 5% CO 2 incubator.

Techniques: Activation Assay, Incubation, Western Blot, Expressing, Inhibition, Activity Assay, Negative Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: GSK-3Beta-Dependent Activation of GEF-H1/ROCK Signaling Promotes LPS-Induced Lung Vascular Endothelial Barrier Dysfunction and Acute Lung Injury

doi: 10.3389/fcimb.2017.00357

Figure Lengend Snippet: GSK-3beta signaling is involved in LPS-induced HPMECs barrier disruption. The HPMECs were plated on the gold microelectrodes. When HPMECs formed monolayers and reached stable TER values, the SB-216763 (20 μM) was added. After 1 h, the medium or LPS (0.1 μg/ml) was added for another 6 h. The HPMEC monolayers permeability was determined by real-time TER measurement (A) . The results of the 3 h LPS stimulation were represented as a histogram in (B) according to the TER curves. * P < 0.05 vs. negative control. # P < 0.05 vs. corresponding LPS-stimulated group.

Article Snippet: Primary human pulmonary micro-vascular endothelial cells (HPMECs) were obtained ScienCell Research Laboratories and maintained in ScienCell Endothelial Cell Medium in a humidified 37°C, 5% CO 2 incubator.

Techniques: Disruption, Permeability, Negative Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: GSK-3Beta-Dependent Activation of GEF-H1/ROCK Signaling Promotes LPS-Induced Lung Vascular Endothelial Barrier Dysfunction and Acute Lung Injury

doi: 10.3389/fcimb.2017.00357

Figure Lengend Snippet: LPS induces degradation of beta-catenin and ZO-1 in HPMECs monolayer. LPS (0.1 μg/ml) induced down-regulation of ZO-1 expression and increase of phosphorylated degradation of beta-catenin in a time-dependent manner (A) . The Western blotting results are presented as a histogram showing the band intensity values (B) . * P < 0.05 vs. LPS un-treatment group.

Article Snippet: Primary human pulmonary micro-vascular endothelial cells (HPMECs) were obtained ScienCell Research Laboratories and maintained in ScienCell Endothelial Cell Medium in a humidified 37°C, 5% CO 2 incubator.

Techniques: Expressing, Western Blot

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: GSK-3Beta-Dependent Activation of GEF-H1/ROCK Signaling Promotes LPS-Induced Lung Vascular Endothelial Barrier Dysfunction and Acute Lung Injury

doi: 10.3389/fcimb.2017.00357

Figure Lengend Snippet: GSK-3beta/GEF-H1/ROCK signaling is required for LPS-induced degradation of beta-catenin and ZO-1. After transfection with GEF-H1 siRNA and Control siRNA for 48 h, HPMECs were treated with SB-216763 (20 μM) and/or Y-27632 (10 μM) for another 1 h prior to LPS stimulation (0.1 μg/ml) for 3 h. The expression of ZO-1 was determined by immunoblotting, and GAPDH protein was used as loading control (A) . The Western blotting results are presented as a histogram showing the band intensity values (B) . The expression of P-beta-catenin was determined by immunoblotting, and GSK-3beta and GAPDH proteins were used as control (C) . The Western blotting results are presented as a histogram showing the band intensity values (D) . * P < 0.05 vs. negative control. # P < 0.05 vs. corresponding LPS-stimulated group. NS, no significance.

Article Snippet: Primary human pulmonary micro-vascular endothelial cells (HPMECs) were obtained ScienCell Research Laboratories and maintained in ScienCell Endothelial Cell Medium in a humidified 37°C, 5% CO 2 incubator.

Techniques: Transfection, Control, Expressing, Western Blot, Negative Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: GSK-3Beta-Dependent Activation of GEF-H1/ROCK Signaling Promotes LPS-Induced Lung Vascular Endothelial Barrier Dysfunction and Acute Lung Injury

doi: 10.3389/fcimb.2017.00357

Figure Lengend Snippet: GSK-3beta/GEF-H1/ROCK pathway is involved in LPS-induced HPMECs barrier disruption by beta-catenin and ZO-1. HPMECs monolayer was pretreated with SB-216763 (20 μM) (A) , GEF-H1 siRNA (B) , or Y-27632 (10 μM) (C) , for indicated times and then was exposed to LPS (0.1 μg/ml) for 3 h before fixation and staining with anti-beta-catenin and anti-ZO-1 antibody as described in Materials and Methods. Beta-catenin (green) and ZO-1 (green) were visualized by immunofluorescence microscopy. Red arrows not only represent the expression of beta-catenin and ZO-1 in the membrane of HPMECs but also represent the cell-cell gaps formation in the ECs monolayer.

Article Snippet: Primary human pulmonary micro-vascular endothelial cells (HPMECs) were obtained ScienCell Research Laboratories and maintained in ScienCell Endothelial Cell Medium in a humidified 37°C, 5% CO 2 incubator.

Techniques: Disruption, Staining, Immunofluorescence, Microscopy, Expressing, Membrane

Journal: Acta biomaterialia

Article Title: Probing prodrug metabolism and reciprocal toxicity with an integrated and humanized multi-tissue organ-on-a-chip platform

doi: 10.1016/j.actbio.2020.02.015

Figure Lengend Snippet: A summary of each organoid or tissue construct type with the cells making them up. Also described are the various cell sources and cell types (primary, cell line, iPSC-derived, etc.), relative percentage in each model, and documented tissue specific functionalities.

Article Snippet: Organoid or tissue construct type Cell types cell source Type of cell Percentage in model Documented functionality Citation Liver Hepatocytes Bioreclamation Primary human 75% Albumin and urea secretion; Drug metabolism; Toxicity responses [ 18 , 47 ] Stellate cells Bioreclamation Primary human 10% Kupffer cells Sekisui XenoTech Primary human 10% Liver-Derived endothelial cells Lonza primary human 5% Cardiac Cardiomyocytes Ncardia iPSC 75% Spontaneous beating; Beating changes in response to drugs; Toxicity responses [ 18 , 47 ] Cardiac fibroblasts ScienCell Primary HUMAN 20% Cardiac endothelial cells ScienCell Primary human 5% Lung (3-tissue system) A549 Alveolar epithelial cells Lonza Human cell line 100% n/a – Lung (6-tissue system) Lung fibroblasts Lonza Primary human 20% n/a – Human bronchial epithelial cells Lonza Primary human 80% Endothelial HUVEC Lonza Primary human 100% n/a – Testis Spermatogonial stem cells Donor via NDRI Primary human ~80% Androgen production; Supporting germ cell differentiation; Toxicity responses; Virus replication [ 34 – 36 ] Leydig cells Donor via NDRI Primary human ~10% Sertoli cells Donor via NDRI Primary human ~10% Peritubular cells Donor via NDRI Primary human Contaminant of SSC populations Brain Brain microvascular endothelial cells Cell Systems Primary human 30% Barrier function to the middle of the spheroid [ 37 ] Brain vascular pericytes ScienCell Primary human 15% Astrocytes ScienCell Primary human 15% Microglia Tempo Bioscience IPSC 5% Oligodendrocytes Tempo Bioscience Oligodendrocyte progenitor cells 15% Neural cells Axol Biosciences Neural stem cells 20% Open in a separate window A summary of each organoid or tissue construct type with the cells making them up.

Techniques: Construct, Cell Differentiation, Virus

Journal: Acta biomaterialia

Article Title: Probing prodrug metabolism and reciprocal toxicity with an integrated and humanized multi-tissue organ-on-a-chip platform

doi: 10.1016/j.actbio.2020.02.015

Figure Lengend Snippet: Drug toxicity assessment of ifosfamide in a 6-organoid system. L/D imaging of liver, cardiac, lung, endothelial, brain, and testis organoids under Control (no drug, a–c), Condition 1 (with ifosfamide and liver organoid, d–f), and Condition 2 (with ifosfamide and without liver organoid, g and h). In Condition 1, the metabolized drug caused downstream toxicity in the brain organoid; in Condition 2, no significant toxicity occurred. Green stain: calcein AM-stained viable cells; Red stain: ethidium homodimer 1-stained dead cells. Scale bar represents 100 μM. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Organoid or tissue construct type Cell types cell source Type of cell Percentage in model Documented functionality Citation Liver Hepatocytes Bioreclamation Primary human 75% Albumin and urea secretion; Drug metabolism; Toxicity responses [ 18 , 47 ] Stellate cells Bioreclamation Primary human 10% Kupffer cells Sekisui XenoTech Primary human 10% Liver-Derived endothelial cells Lonza primary human 5% Cardiac Cardiomyocytes Ncardia iPSC 75% Spontaneous beating; Beating changes in response to drugs; Toxicity responses [ 18 , 47 ] Cardiac fibroblasts ScienCell Primary HUMAN 20% Cardiac endothelial cells ScienCell Primary human 5% Lung (3-tissue system) A549 Alveolar epithelial cells Lonza Human cell line 100% n/a – Lung (6-tissue system) Lung fibroblasts Lonza Primary human 20% n/a – Human bronchial epithelial cells Lonza Primary human 80% Endothelial HUVEC Lonza Primary human 100% n/a – Testis Spermatogonial stem cells Donor via NDRI Primary human ~80% Androgen production; Supporting germ cell differentiation; Toxicity responses; Virus replication [ 34 – 36 ] Leydig cells Donor via NDRI Primary human ~10% Sertoli cells Donor via NDRI Primary human ~10% Peritubular cells Donor via NDRI Primary human Contaminant of SSC populations Brain Brain microvascular endothelial cells Cell Systems Primary human 30% Barrier function to the middle of the spheroid [ 37 ] Brain vascular pericytes ScienCell Primary human 15% Astrocytes ScienCell Primary human 15% Microglia Tempo Bioscience IPSC 5% Oligodendrocytes Tempo Bioscience Oligodendrocyte progenitor cells 15% Neural cells Axol Biosciences Neural stem cells 20% Open in a separate window A summary of each organoid or tissue construct type with the cells making them up.

Techniques: Imaging, Staining

Journal: Acta biomaterialia

Article Title: Probing prodrug metabolism and reciprocal toxicity with an integrated and humanized multi-tissue organ-on-a-chip platform

doi: 10.1016/j.actbio.2020.02.015

Figure Lengend Snippet: Viability quantification under insult by ifosfamide in a 6-organoid system. Quantification of live cell ratios (L/L + D) for liver, cardiac, lung, endothelial, brain, and testis organoids under Control (no drug), Condition 1 (drug with liver organoid), and Condition 2 (drug without liver organoid). A significant reduction in viability is observed for the brain organoid under Condition 1 (red). Statistical significance: ** p < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Organoid or tissue construct type Cell types cell source Type of cell Percentage in model Documented functionality Citation Liver Hepatocytes Bioreclamation Primary human 75% Albumin and urea secretion; Drug metabolism; Toxicity responses [ 18 , 47 ] Stellate cells Bioreclamation Primary human 10% Kupffer cells Sekisui XenoTech Primary human 10% Liver-Derived endothelial cells Lonza primary human 5% Cardiac Cardiomyocytes Ncardia iPSC 75% Spontaneous beating; Beating changes in response to drugs; Toxicity responses [ 18 , 47 ] Cardiac fibroblasts ScienCell Primary HUMAN 20% Cardiac endothelial cells ScienCell Primary human 5% Lung (3-tissue system) A549 Alveolar epithelial cells Lonza Human cell line 100% n/a – Lung (6-tissue system) Lung fibroblasts Lonza Primary human 20% n/a – Human bronchial epithelial cells Lonza Primary human 80% Endothelial HUVEC Lonza Primary human 100% n/a – Testis Spermatogonial stem cells Donor via NDRI Primary human ~80% Androgen production; Supporting germ cell differentiation; Toxicity responses; Virus replication [ 34 – 36 ] Leydig cells Donor via NDRI Primary human ~10% Sertoli cells Donor via NDRI Primary human ~10% Peritubular cells Donor via NDRI Primary human Contaminant of SSC populations Brain Brain microvascular endothelial cells Cell Systems Primary human 30% Barrier function to the middle of the spheroid [ 37 ] Brain vascular pericytes ScienCell Primary human 15% Astrocytes ScienCell Primary human 15% Microglia Tempo Bioscience IPSC 5% Oligodendrocytes Tempo Bioscience Oligodendrocyte progenitor cells 15% Neural cells Axol Biosciences Neural stem cells 20% Open in a separate window A summary of each organoid or tissue construct type with the cells making them up.

Techniques:

Journal: Journal of Diabetes Investigation

Article Title: Long non‐coding ribonucleic acid ATP2B1‐AS1 modulates endothelial permeability through regulating the miR‐4729–IQGAP2 axis in diabetic retinopathy

doi: 10.1111/jdi.13740

Figure Lengend Snippet: Identification of ATPase plasma membrane Ca 2+ transporting 1 antisense ribonucleic acid 1 (ATP2B1‐AS1) in diabetic retinopathy (DR) and high‐glucose‐treated high‐glucose‐treated human retinal endothelial cells (HRECs). (a) The heatmap of the differentially expressed genes in low glucose (LG) and high glucose (HG). Upregulated genes and downregulated genes are shown in red and blue. (b) Volcano plots showing long non‐coding ribonucleic acids expression in the LG and HG groups. The red dots show the significant expressed genes. (c) Reverse transcription quantitative polymerase chain reaction was carried out to detect ATP2B1‐AS1 levels in 5 mmol/L or 25 mmol/L glucose treated HRECs. (d) Reverse transcription quantitative polymerase was carried out to distinguish the level of ATP2B1‐AS1 in blood samples obtained from DR patients ( n = 30) and healthy individuals. All values were represented by the mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human retinal endothelial cells (HRECs) and 293T cells were purchased from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Clinical Proteomics, Membrane, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Journal of Diabetes Investigation

Article Title: Long non‐coding ribonucleic acid ATP2B1‐AS1 modulates endothelial permeability through regulating the miR‐4729–IQGAP2 axis in diabetic retinopathy

doi: 10.1111/jdi.13740

Figure Lengend Snippet: ATPase plasma membrane Ca 2+ transporting 1 antisense ribonucleic acid 1 (ATP2B1‐AS1) prevents cell proliferation, migration, angiogenesis and permeability. (a) Reverse transcription quantitative polymerase chain reaction was made to measure the expression of ATP2B1‐AS1 after transfecting plasmid cloning deoxyribonucleic acid (pcDNA)‐long non‐coding ribonucleic acids (lncRNA) ATP2B1‐AS1 (pcDNA‐lnc) and (short hairpin RNA‐lncRNA ATP2B1‐AS1; shR‐lnc) into high‐glucose‐treated human retinal endothelial cells (HRECs). (b) The level of ATP2B1‐AS1 was detected by reverse transcription polymerase chain reaction after transfecting pcDNA‐lnc and shR‐lnc into HRECs by Cell Counting Kit‐8 assay. (c) Proliferation of HRECs was detected by Cell Counting Kit‐8 assay. (d, e) Migration ability was measured by wound healing migration assay and transwell assay. (f) Tube formation assay was used to distinguish angiogenesis ability in HRECs. (g) Cell junctional assembly formation of CDH5 staining. (h) Vascular permeability was detected by using evans blue injection. All values were represented by the mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human retinal endothelial cells (HRECs) and 293T cells were purchased from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Clinical Proteomics, Membrane, Migration, Permeability, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Plasmid Preparation, Cloning, shRNA, Polymerase Chain Reaction, Cell Counting, Transwell Assay, Tube Formation Assay, Staining, Injection, Standard Deviation

Journal: Journal of Diabetes Investigation

Article Title: Long non‐coding ribonucleic acid ATP2B1‐AS1 modulates endothelial permeability through regulating the miR‐4729–IQGAP2 axis in diabetic retinopathy

doi: 10.1111/jdi.13740

Figure Lengend Snippet: ATPase plasma membrane Ca 2+ transporting 1 antisense ribonucleic acid 1 (ATP2B1‐AS1) sponges microRNA (miR)‐4729. The microRNAs lists and scores on predicted by using the MicroRNA Target Prediction Database. (b) Predicted miR‐4729 binding sites in 3′UTR of ATP2B1‐AS1 and dual luciferase report assay in ATP2B1‐AS1‐wild type (WT) or ATP2B1‐AS1‐mutation (MUT) co‐transfected with miR negative control (NC) or miR‐4729 mimics. (c) Level of miR‐4729 in high‐glucose‐treated human retinal endothelial cells (HRECs) transfected with shR‐lnc or pcDNA‐lnc. (d) miR‐4729 expression in blood from diabetes retinopathy (DR) patients ( n = 30) and non‐DR individuals. (e) Pearson's correlation analysis was used to check the relationship between ATP2B1‐AS1 and miR‐4729. All values were represented by the mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human retinal endothelial cells (HRECs) and 293T cells were purchased from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Clinical Proteomics, Membrane, Binding Assay, Luciferase, Mutagenesis, Transfection, Negative Control, Expressing, Standard Deviation

Journal: Journal of Diabetes Investigation

Article Title: Long non‐coding ribonucleic acid ATP2B1‐AS1 modulates endothelial permeability through regulating the miR‐4729–IQGAP2 axis in diabetic retinopathy

doi: 10.1111/jdi.13740

Figure Lengend Snippet: ATPase plasma membrane Ca 2+ transporting 1 antisense ribonucleic acid 1 (ATP2B1‐AS1) reduced high glucose‐treated high‐glucose‐treated human retinal endothelial cells (HRECs) cell proliferation, migration, angiogenesis and permeability through regulating microRNA (miR)‐4729–IQ motif‐containing GTPase‐activating protein 2 (IQGAP2) axis. (a) Schematic indicating the miR‐4729 sites in IQGAP2 and dual luciferase assay in IQGAP2‐wild type (WT) or IQGAP2‐mutation (MUT) treated HRECs co‐transfected with miR‐NC or miR‐4729 mimics. (b) The protein IQGAP2 level was detected by WB after transfection. (c) HRECs proliferation was detected by Cell Counting Kit‐8 assay after transfection. (d, e) Migration ability was measured by wound healing migration assay and transwell assay after transfection. (f) Tube formation assay was used to detect the ability of angiogenesis in HRECs after transfection. (g) Cell junctional assembly formation of VE‐cadherin staining after transfection. All values were represented by the mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human retinal endothelial cells (HRECs) and 293T cells were purchased from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Clinical Proteomics, Membrane, Migration, Permeability, Luciferase, Mutagenesis, Transfection, Cell Counting, Transwell Assay, Tube Formation Assay, Staining, Standard Deviation