Journal: Cell reports

Article Title: Cancer-associated fibroblasts maintain critical pancreatic cancer cell lipid homeostasis in the tumor microenvironment

doi: 10.1016/j.celrep.2024.114972

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Pancreatic Microvascular Endothelial Cells were purchased from iXCell Biotechnologies and cultured in Endothelial Cell Growth Media (iXCell, MD-0010).

Techniques: Recombinant, Western Blot, Transfection, Reverse Transcription, Flow Cytometry, Bicinchoninic Acid Protein Assay, Plasmid Preparation, ROS Assay, Mass Spectrometry, shRNA, Control, Software, Microscopy, Real-time Polymerase Chain Reaction

Journal: Bioscience, biotechnology, and biochemistry

Article Title: Inhibition of miR-21-5p suppresses high glucose-induced proliferation and angiogenesis of human retinal microvascular endothelial cells by the regulation of AKT and ERK pathways via maspin.

doi: 10.1080/09168451.2018.1459179

Figure Lengend Snippet: Figure 1. Effects of high glucose on cell proliferation and miR-21-5p, VEGF, VEGFR2 expression of HRMECs. Cell proliferation was measured by MTT assay (a). The level of miR-21-5p was detected by real-time PCR (b). The mRNA levels of VEGF (c) and VEGFR2 (e) were measured by real-time PCR. The protein levels of VEGF (d) and VEGFR2 (f) were determined by western blot. Experiments were repeated at least for three times. **p<0.05, **p<0.01 vs. control cells, ##p<0.01 vs. 24 h treated cells.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) were purchased from Angio-Proteomie (Boston, MA, USA) and cultured in endothelial cell medium (ECM, Sciencell) supplemented with 5% fetal bovine serum (FBS, Sciencell), 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C under 5% CO2 and 95% ambient air.

Techniques: Expressing, MTT Assay, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Bioscience, biotechnology, and biochemistry

Article Title: Inhibition of miR-21-5p suppresses high glucose-induced proliferation and angiogenesis of human retinal microvascular endothelial cells by the regulation of AKT and ERK pathways via maspin.

doi: 10.1080/09168451.2018.1459179

Figure Lengend Snippet: Figure 2. Effects of miR-21-5p on the proliferation of HRMECs. Cell proliferation was detected by MTT assay (a). Expression of Ki67 was measured by immunofluorescence (b, c). Typical images from three repeats were shown. **p<0.05, **p<0.01 vs. control group, #p<0.05, ##p<0.01 vs. high glucose + NC inhibitor-treated cells. NC: negative control.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) were purchased from Angio-Proteomie (Boston, MA, USA) and cultured in endothelial cell medium (ECM, Sciencell) supplemented with 5% fetal bovine serum (FBS, Sciencell), 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C under 5% CO2 and 95% ambient air.

Techniques: MTT Assay, Expressing, Immunofluorescence, Control, Negative Control

Journal: Bioscience, biotechnology, and biochemistry

Article Title: Inhibition of miR-21-5p suppresses high glucose-induced proliferation and angiogenesis of human retinal microvascular endothelial cells by the regulation of AKT and ERK pathways via maspin.

doi: 10.1080/09168451.2018.1459179

Figure Lengend Snippet: Figure 3. Effects of miR-21-5p on the angiogenesis of HRMECs. Cell migration was detected by wound healing assay (a, b). Expression of MMP-2 and MMP-9 were measured by western blot (c). Tube formation and branching points were determined (d, e). Expression of HIF-1α and VEGF were measured by western blot (f). Photographs are representative of at least three individual experiments. **p < 0.01 vs. control cells, #p < 0.05, ##p < 0.01 vs. high glucose + NC inhibitor-treated cells. NC: negative control.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) were purchased from Angio-Proteomie (Boston, MA, USA) and cultured in endothelial cell medium (ECM, Sciencell) supplemented with 5% fetal bovine serum (FBS, Sciencell), 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C under 5% CO2 and 95% ambient air.

Techniques: Migration, Wound Healing Assay, Expressing, Western Blot, Control, Negative Control

Journal: Bioscience, biotechnology, and biochemistry

Article Title: Inhibition of miR-21-5p suppresses high glucose-induced proliferation and angiogenesis of human retinal microvascular endothelial cells by the regulation of AKT and ERK pathways via maspin.

doi: 10.1080/09168451.2018.1459179

Figure Lengend Snippet: Figure 5. Effects of PI3K/AKT and ERK on the angiogenesis of HRMECs. Cell migration was detected by wound healing assay (a, b). Expression of MMP-2 and MMP-9 were measured by western blot (c). Tube formation and branching points were determined (d, e). Expression of HIF-1α and VEGF were measured by western blot (f). Photographs are representative of three individual experiments. **p < 0.05, **p < 0.01 vs. high glucose-treated cells.

Article Snippet: Human retinal microvascular endothelial cells (HRMECs) were purchased from Angio-Proteomie (Boston, MA, USA) and cultured in endothelial cell medium (ECM, Sciencell) supplemented with 5% fetal bovine serum (FBS, Sciencell), 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C under 5% CO2 and 95% ambient air.

Techniques: Migration, Wound Healing Assay, Expressing, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: CLIC5A binds to and stabilizes the open and active conformation of ezrin

doi: 10.1016/j.jbc.2025.110646

Figure Lengend Snippet: ERM and Rac-1 activation are amplified by the CLIC5A/ezrin interaction. A , endogenous pERM abundance in lysates and detergent resistant pellets of COS-7 cells transfected with GFP-CLIC5A cDNA and increasing concentrations of GFP-ezrin 432-586 (T567D) cDNA . GFP-CLIC5A cDNA was kept constant and the GFP-CLIC5A: GFP-ezrin 432-586 (T567D) cDNA transfection ratio was 1:1, 1:2, 1:4, and 1:8. Left panel , representative WB. Right panel, densitometric quantification of the endogenous pERM: endogenous ezrin ratio (mean ± SD, n = 3 independent experiments, one-way ANOVA: F = 7.90; p = 0.0007. p values shown represent post hoc Dunnett’s multiple comparisons). B , pERM abundance in lysates and detergent resistant pellets of COS-7 cells transiently transfected with GFP-CLIC5A cDNA with or without an 8-fold excess of GFP- e zrin 432-570 cDNA. Left panel , representative WB. Right panel, densitometric quantification of the endogenous pERM: endogenous ezrin ratio (mean ± S.D., n = 3 biologically independent experiments, two-way ANOVA: interaction F = 12.25; p = 0.01; CLIC5A effect: F = 39.63, p = 0.0002; ezrin 432-570 effect F = 22.58. p = 0.0014; p values shown represent post hoc Tukey’s multiple comparisons). C , endogenous Rac1 WB for COS-7 cell lysates and endogenous Rac1-GTP captured by PAK-PBD pulldown (PD) from COS-7 cells transiently transfected with GFP-CLIC5A cDNA with or without an 8-fold excess of GFP-ezrin 432-586 (T567D) cDNA. Left panel, representative WB. Right panel: densitometric quantification of endogenous Rac1-GTP/total endogenous Rac1 (mean ± SD, n = 3 independent experiments, two-way ANOVA: interaction F = 6.32, p = 0.036; CLIC5A effect F = 29.25, p = 0.001; ezrin 432–586 effect F = 5.88; p = 0.042, p values for post hoc Tukey’s multiple comparisons are shown). D , PAK-PBD pulldown (PD) of endogenous Rac1-GTP from lysates of COS-7 cells transfected with GFP-CLIC5A cDNA with or without an 8-fold excess of transiently expressed GFP-ezrin 432 - 570 cDNA. Left panel, representative WB. Right panel, densitometric quantification of endogenous Rac1-GTP/total endogenous Rac1 (mean ± S.D., n = 3 independent experiments, two-way ANOVA: interaction F = 0.029, p = 0.87; CLIC5A effect F = 62.17 p < 0.0001; ezrin 432–570 effect F = 0.012; p = 0.91, p values for post hoc Tukey’s multiple comparisons are shown). E , endogenous Rac1-GTP abundance in human glomerular endothelial cells (hGENs) determined by Rac1-GTP G-LISA. The cells were transduced with control adenoviral-vector (ad-Vector) or untagged CLIC5A cDNA in the same vector (ad- CLIC5A ) at an increasing multiplicity of infection (MOI) (mean ± SD, n = 3 independent experiments). F , representative WB of lysates from hGEN cells transduced with ad-Vector or ad- CLIC5A with or without ezrin-specific siRNA. G , change relative to baseline of Rac1-GTP in hGEN cells transduced with 30 MOI ad-Vector or ad- CLIC5A with or without ezrin-specific siRNA (mean ± SD, n = 4 independent experiments, two-way ANOVA: interaction F = 6.35, p = 0.027; CLIC5A effect F = 34.18, p < 0.0001; ezrin siRNA effect F = 6.86; p = 0.023, p values shown represent post hoc Tukey’s multiple comparisons). H , coimmunoprecipitation of endogenous Rho-GDI with endogenous ezrin in the presence and absence of transiently expressed GFP-CLIC5A. Left panel: representative α-Rho GDI, α-ezrin, and α-CLIC5A WB of lysates (input) and α-ezrin immunoprecipitates (IP). Right panel: Quantification of endogenous Rho-GDI and endogenous ezrin immunoprecipitated with α-ezrin antibodies (n = 3 biologically distinct experiments, mean ± SD, Student’s t test). CLIC, chloride intracellular channel; PBD, protein binding domain; pERM, phosphorylated ezrin, radixin, and moesin proteins; Rho-GDI, Rho guanine nucleotide dissociation inhibitor; WB, Western blot.

Article Snippet: Mycoplasma-free primary human glomerular endothelial cells (hGENs) were purchased from Angio-Proteomie (# cAP-0004) and maintained in EGM-2 MV Bulletkit growth media (# CC-3162, Lonza) containing 5% FBS and 1% penicillin/streptomycin at 37 °C in humidified air containing 5% CO 2 .

Techniques: Activation Assay, Amplification, Transfection, Transduction, Control, Plasmid Preparation, Infection, Immunoprecipitation, Protein Binding, Western Blot

Journal: bioRxiv

Article Title: Development of a Synthetic Hydrogel to Foster Microvascularization of an Endometriosis Microphysiological System

doi: 10.1101/2025.10.06.680733

Figure Lengend Snippet: A) Endometriosis lesions establish complex microenvironments consisting of epithelial, stromal, endothelial, and immune cells. B) Droplet hydrogel cultures were used to assess organoid emergence from single cell. C) v-CS-ECM supports organoid emergence from single cell over the course of a week. D) EEOs, HUTMVECs, and ESCs were combined in a hydrogel precursor prior to injection in a microfluidic device and cultured for 6 days. E) V-CS-ECM supports EEO growth within HUTMVEC-ESC networks. Scale Bar: 500μm.

Article Snippet: GFP-labeled Human umbilical vein endothelial cells (HUVECs) and GFP-labeled human uterine microvascular endothelial cells (HUTMVECs) were purchased from Angio-Proteomie (cAP-0001GFP and cAP-0023GFP).

Techniques: Injection, Cell Culture

Journal: Scientific Reports

Article Title: In vitro and in vivo validation of a novel 3D-printed vessel anastomosis device for microvascular surgery

doi: 10.1038/s41598-026-39181-4

Figure Lengend Snippet: Application of microvascular anastomosis in clinical practice. The need to connect blood vessels (anastomosis) is found in many surgical specialities, such as neurosurgery, oral and maxillofacial surgery, cardiothoracic surgery, hand surgery, liver and kidney transplants, vascular surgery and lower limb reconstructive surgery.

Article Snippet: The cytotoxicity test was conducted using human cardiac microvascular endothelial cells (HCMECs) from PromoCell (PromoCell GmbH, Heidelberg.

Techniques:

Journal: bioRxiv

Article Title: Vascularized tumor organoids enable immunotherapy testing in tumor-remodeled stroma

doi: 10.64898/2026.01.23.701300

Figure Lengend Snippet: a, Endothelial activation by TNFα induces robust up-regulation of adhesion molecules across the microvascular network, shown as intensity heat maps for ICAM1 and VCAM1 (Control vs TNFα). b, Representative confocal images of the vascularized channel showing endothelial cells (cyan) and PBMCs (magenta/red). TNFα priming increases leukocyte tethering/adhesion and transendothelial migration (white dashed outlines; higher-magnification insets). c, Quantification of leukocyte–vessel interactions per chip, partitioned into adherent (grey) and extravasated (magenta) cells; TNFα markedly elevates both behaviors (bars = mean, error bars = s.d.; points = individual chips). d, Secreted CXCL10 measured from chip effluents rises after TNFα stimulation (points = chips; bars = mean ± s.d.). Data are representative of independent chips per condition.

Article Snippet: Human liver sinusoidal microvascular endothelial cells (LSMVECs) (Angio-Proteomie) expressing GFP were cultured in EGM-2 MV (Lonza) up to passage 9.

Techniques: Activation Assay, Control, Migration

Journal: bioRxiv

Article Title: Vascularized tumor organoids enable immunotherapy testing in tumor-remodeled stroma

doi: 10.64898/2026.01.23.701300

Figure Lengend Snippet: a, Time-lapse montage of a microvascular network co-cultured with healthy colon organoids during perfusion of fluorescent 70-kDa dextran (pseudocolor; yellow/white = higher intensity) at 0, 100, 200 and 300 s,showing rapid intraluminal filling adjacent to organoids. b, Wide-field views of the perfused channel along the chip with (top) and without (bottom) organoids illustrate uniform tracer distribution throughout the network. Images are representative of independent chips.

Article Snippet: Human liver sinusoidal microvascular endothelial cells (LSMVECs) (Angio-Proteomie) expressing GFP were cultured in EGM-2 MV (Lonza) up to passage 9.

Techniques: Cell Culture

Journal: Cellular and Molecular Immunology

Article Title: Arterial thrombosis in the context of HCV-associated vascular disease can be prevented by protein C

doi: 10.1038/cmi.2016.10

Figure Lengend Snippet: Effect of poly (I:C) on the endothelial expression of procoagulatory factors and clotting time. HMEC were stimulated with poly (I:C) (10 μg/ml) for 12 h and the expression of tissue factor (a) and PAI-1 (b) was analyzed by RT–PCR (n=4, *P<0.05. mean±s.e., statistics with t-test (sigma plot); rel. to ct, relative to control). Comparable results were obtained in two series of independent experiments. (c) HMEC were stimulated with poly (I:C) (10 μg/ml) or TNFα (5 ng/ml) as a positive control for 24 h and then lysed. Whole blood samples were stimulated with cell lysates and the clotting time was analyzed as described in the Materials and methods section (n=5–6, *P<0.05. mean±s.e., statistics with t-test (sigma plot); rel. to ct, relative to control). Comparable results were obtained in two series of independent experiments. HMEC, human microvascular endothelial cell; poly (I:C), polyriboinosinic:polyribocytidylic acid; RT–PCR, reverse transcription–PCR. **P<0.01.

Article Snippet: Human microvascular endothelial cells (HMEC) were provided by Ades et al. 14 and were cultured in M199 media supplement with 10% fetal calf serum, 10% endothelial growth media (PromoCell, Heidelberg, Germany) and 1% penicillin/streptomycin as described previously.

Techniques: Expressing, Coagulation, Reverse Transcription Polymerase Chain Reaction, Positive Control

Journal: Cellular and Molecular Immunology

Article Title: Arterial thrombosis in the context of HCV-associated vascular disease can be prevented by protein C

doi: 10.1038/cmi.2016.10

Figure Lengend Snippet: Poly (I:C) did not influence platelet aggregation and activation. Light transmission aggregometry (the method by Born)18 was performed in platelet-rich-plasma (PRP) from healthy human volunteers, as described in the Materials and methods section. The percent light transmission of platelet-rich plasma (PRP) was compared with platelet poor plasma (PPP) on stimulation with poly (I:C) (10 μg/ml) or ADP (10 μM) (a and b). PRP was incubated with poly (I:C) (10 μg/ml) for different time intervals (10, 20, 30, 45 min) and ADP-dependent (5 μM) platelet aggregation was analyzed (c). PRP was incubated with poly (I:C) (10 μg/ml) for 15 min in the presence of ADP at a low concentration (5 μM), ADP at a high concentration (10 μM), thrombin-receptor-activated peptide (TRAP) (20 μM) or collagen (10 μg/ml) and platelet aggregation was analyzed (n=4, P>0.05, mean±s.e., statistics with t-test (sigma plot)) Comparable results were obtained in two series of independent experiments. (d) Human platelets were isolated as described in the Materials and methods section. The platelets were stimulated with poly (I:C) (10 μg/ml) for different time intervals (10, 60 min) alone or in the presence of thrombin (2 U/ml) and FACS analysis with a monoclonal antibody against P-selectin (e and f) and GPIIbIIIa (g and h) was performed (n=3–4, P>0.05, mean±s.e., statistics with one-way ANOVA (sigma plot)). Comparable results were obtained in two series of independent experiments. ANOVA, analysis of variance; HMEC, human microvascular endothelial cell; poly (I:C), polyriboinosinic:polyribocytidylic acid.

Article Snippet: Human microvascular endothelial cells (HMEC) were provided by Ades et al. 14 and were cultured in M199 media supplement with 10% fetal calf serum, 10% endothelial growth media (PromoCell, Heidelberg, Germany) and 1% penicillin/streptomycin as described previously.

Techniques: Activation Assay, Transmission Assay, Incubation, Concentration Assay, Isolation

Journal: bioRxiv

Article Title: The pro-inflammatory response to influenza A virus infection is fueled by endothelial cells

doi: 10.1101/2022.08.19.504520

Figure Lengend Snippet: To evaluate replication efficiency, lung microvascular endothelial cells (LMECs) were plated on the (A) apical or (B) basolateral side of a transwell filter. LMECs were inoculated with pH1N1, H1N1, or H3N2 virus at MOI 1 and at the indicated time points supernatants of the apical as well as basolateral side were harvested and virus titers were determined by endpoint titration. Infection efficiency was determined by immunofluorescence staining. LMECs plated on the (C) apical and (D) basolateral were inoculated with pH1N1, H1N1, or H3N2 virus. Cells were fixed 24 hours post inoculation and stained for the endothelial cell marker Vascular Endothelial-Cadherin (VE-CAD, magenta) and influenza A virus nucleoprotein (NP (green). Hoechst (blue) was used to visualize nuclei. (E) Percentage of infection determined by flow cytometry at 24 and 72 hours post inoculation. (F) Viral RNA genome copies were quantified by quantitative real time PCR at indicated time points. Data represent mean +/- standard deviation (SD) from at least three independent experiments performed in biological duplicates and flow cytometry was performed in biological triplicates. A one-way ANOVA multiple comparison test was used to compare groups (*< 0.05, **<0.01, ***<0.005). Scale bar: 20 μm.

Article Snippet: Primary human lung microvascular endothelial cells (LMEC) were purchased at passage 3 from PromoCell-PromoKine (#C-12285) and cultured in 1% gelatine coated cell cultures flasks with Endothelial Cell Growth Medium MV-2 kit according to the manufacturer’s protocol (#C-22121, PromoCell-PromoKine), LMECs were only used up to passage 10 to ensure organ specificity.

Techniques: Titration, Infection, Immunofluorescence, Staining, Marker, Flow Cytometry, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: bioRxiv

Article Title: The pro-inflammatory response to influenza A virus infection is fueled by endothelial cells

doi: 10.1101/2022.08.19.504520

Figure Lengend Snippet: (A) Well-differentiated airway organoids at air-liquid interface (AO at ALI) in co-culture with lung microvascular endothelial cells (LMECs) were inoculated with pH1N1, H1N1 or H3N2 virus at MOI 1. At the indicated timepoints virus titers were determined in the supernatants of the apical and basolateral compartments. (B) Detection of influenza A virus (IAV) nucleoprotein (NP) by immunohistochemistry of the AO at ALI-LMEC co-cultures 24 hours post inoculation (C) Hematoxylin and eosin (H&E) staining of the co-cultures 72 hours post inoculation (scale bar 20 μm) (D) At 72 hours post inoculation well-differentiated AO at ALI were stained for IAV NP (green), the cilia marker acetylated-α-tubulin (cyan) and the tight-junction marker Zona occludin 1 (ZO-1, magenta) on the apical compartment of the transwell. The basolateral compartment containing the LMECs was stained for IAV NP (green) and the endothelial cell marker Vascular-Endothelial Cadherin (VE-CAD, magenta). In both cases the nuclei were visualized with Hoechst (scale bar 20 μm). (E) Epithelial cells (AO at ALI) or endothelial-epithelial co-cultures were inoculated with pH1N1, H1N1 or H3N2 virus at MOI 1. At 24 hours post-inoculation cytokines were measured in the apical compartment using the Legendplex assay. Data represented here show individual data points of cytokines derived from three independent experiments performed in biological duplicates and the mean +/- standard deviation (SD) is depicted. Mock of each condition was subtracted from the values of virus infected cells. Statistical significance was determined with Students-T-test (*<0.05, **<0.01, ***<0.005, ****<0.001).

Article Snippet: Primary human lung microvascular endothelial cells (LMEC) were purchased at passage 3 from PromoCell-PromoKine (#C-12285) and cultured in 1% gelatine coated cell cultures flasks with Endothelial Cell Growth Medium MV-2 kit according to the manufacturer’s protocol (#C-22121, PromoCell-PromoKine), LMECs were only used up to passage 10 to ensure organ specificity.

Techniques: Co-Culture Assay, Immunohistochemistry, Staining, Marker, Derivative Assay, Standard Deviation, Infection

Journal: bioRxiv

Article Title: The pro-inflammatory response to influenza A virus infection is fueled by endothelial cells

doi: 10.1101/2022.08.19.504520

Figure Lengend Snippet: Lung microvascular endothelial cells (LMECs) or differentiated airway organoids at air-liquid interface in co-culture with LMECs were inoculated with pH1N1 virus at MOI 1. (A) percentage of infection and (B) viral genome copies in LMEC single cultures compared to co-cultures were determined by flow cytometry or qRT-PCR at 24 hours post-inoculation. Data represented here show pooled data of virus titers derived from three independent experiments performed in biological duplicates and the mean +/- standard deviation is depicted. A student T-test was used to compare groups (*<0.05, **<0.01, ***<0.005, ****<0.001).

Article Snippet: Primary human lung microvascular endothelial cells (LMEC) were purchased at passage 3 from PromoCell-PromoKine (#C-12285) and cultured in 1% gelatine coated cell cultures flasks with Endothelial Cell Growth Medium MV-2 kit according to the manufacturer’s protocol (#C-22121, PromoCell-PromoKine), LMECs were only used up to passage 10 to ensure organ specificity.

Techniques: Co-Culture Assay, Infection, Flow Cytometry, Quantitative RT-PCR, Derivative Assay, Standard Deviation

Journal: bioRxiv

Article Title: The pro-inflammatory response to influenza A virus infection is fueled by endothelial cells

doi: 10.1101/2022.08.19.504520

Figure Lengend Snippet: (A) Endothelial cells (lung microvascular endothelial cells), epithelial cells (airway organoids at air-liquid interface) and endothelial-epithelial co-cultures were inoculated with pH1N1, H1N1 and H3N2 virus at MOI 1. At 24 hours post-inoculation cytokines were measured in the basolateral compartment using a Legendplex assay. Data represented here show individual data points of cytokines derived from three independent experiments performed in biological duplicates and the mean +/- standard deviation is depicted. Mock of each condition was subtracted from the values of virus inoculated cultures. Statistical significance was determined with One-Way Anova and each group was compared to each other (*<0.05, **<0.01, ***<0.005, ****<0.001).

Article Snippet: Primary human lung microvascular endothelial cells (LMEC) were purchased at passage 3 from PromoCell-PromoKine (#C-12285) and cultured in 1% gelatine coated cell cultures flasks with Endothelial Cell Growth Medium MV-2 kit according to the manufacturer’s protocol (#C-22121, PromoCell-PromoKine), LMECs were only used up to passage 10 to ensure organ specificity.

Techniques: Derivative Assay, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Modular Microphysiological System for Modeling of Biologic Barrier Function

doi: 10.3389/fbioe.2020.581163

Figure Lengend Snippet: Recapitulating alveolar microenvironment: (A) Cartoon of alveolar expansion during inspiration. Created with BioRender.com. (B) Calculated bi-axial strain in response to applied pressure in the microfluidic chip. (C) Application of a cyclic pressure in the apical channel using a sine wave to mimic breathing at approximately 20 breaths per minute. A constant pressure was applied to the basal channel to drive the flow of PBS. (D) Workflow for development of ALI co-culture. Endothelial cells (LMECs) were first seeded on the basal surface of a transwell culture insert, followed alveolar epithelial cells (AECs). Dexamethasone was added to the apical chamber on Day 3 to enhance the epithelial cell barrier. On Day 7, the ALI was induced by removing media from the apical chamber. On Day 10, the membrane was cut from the transwell support and bonded into the microfluidic chip and exposed to dynamic strain. (E) TEER measurements during liquid-liquid co-culture in the transwell with (Dex +) and without (Dex-) dexamethasone. Statistical analysis by unpaired t -test ( N = 3). (F) 3D visualization of fluorescent z -stack after culture for 24 h under cyclic pressure exposure in the MPS. Cytokeratin (green) staining shows the alveolar endothelial cells on the apical side of the membrane, while PECAM1 (red) staining indicates the lung microvascular endothelial cells on the basal side.

Article Snippet: Primary human glomerular microvascular endothelial cells (GMECs; Cell Systems) were culture per the manufacturer in complete endothelial cell growth media 2 (PromoCell).

Techniques: Co-Culture Assay, Staining