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thincert chambers for rhe models  (Greiner Bio)


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    Structured Review

    Greiner Bio thincert chambers for rhe models
    Thincert Chambers For Rhe Models, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 95/100, based on 430 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/insertion+model/Thincert+Cell+Culture+Insert+For+12+Well+Plates/pmc12989852-85-0-6
    Average 95 stars, based on 430 article reviews
    thincert chambers for rhe models - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Cell Culture:

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1–2 h incubation. ..

    Article Title: Transcriptomic signatures of IPF in ALI-cultured airway cells and their therapeutic implications.
    Article Snippet: HBECs were cultured following previously published methods.45 Briefly, cells were grown in bronchial expansion medium: DMEM/F12 (Gibco, 21331020) supplemented with Bovine Pituitary Extract (0.004 mL/mL), Epidermal Growth Factor (10 ng/mL), Insulin (5 μg/mL), Hydrocortisone (0.5 μg/mL), Triiodo‐L‐Thyronine (6.7 ng/mL) and Transferrin (10 μg/mL) (PromoCell, C- 39160), 30 nM Sodium Selenite (Merck, S5261), 10 μM Ethanolamine (Merck, E0135), 10 μM Phosphorylethanolamine (Merck, P0503), 0.18 mM Adenine (Merck, A2786), 0.5 μM Sodium Pyruvate (Gibco, 11 360–070), 1X GlutaMAX (Gibco, 35050038), 15 mM HEPES (Gibco, 15 630–056), and in the presence of 10 μM ROCK Inhibitor (StemCell, 72308). .. After reaching confluence, cells were transferred into 12- well ThinCert cell culture inserts with 0.4 μm pores (Greiner Bio- One, 665641). ..

    Article Title: Biodegradable Carbonate Nanogels Loaded with Anti MFAP-5 siRNA for Anti-stromal Therapy of Hepatocellular Carcinoma.
    Article Snippet: .. NIH/3T3 cells (10,000 cells/well) were seeded into 12-well plates, while Dt81Hepa1-6 cells (5,000 cells/insert) were seeded in cell culture inserts (ThinCert, 8 μm pore size, Greiner, Kremsmünster, Austria) and placed in the same wells for coculture. .. After 5 days of co-cultivation at 37 ◦C with 5% CO2 , NG:siMFAP5 was added at final siRNA concentrations of 25, 50, and 100 n m . For positive controls, cells were treated with anti-MFAP5 siRNA using the transfection agent Dharmafect (Dharmacon, Lafayette, USA) at 100 n m siRNA, following the manufacturer’s protocol.

    Article Title: Contact-compression induces inflammatory and remodeling responses in bronchial epithelial cells.
    Article Snippet: 16 Aims and objectives: Lung implantable devices, such as stents and valves are used as treatment for lung 17 cancer and COPD.. They apply continuous compressive stress to airway tissue, potentially triggering 18 adverse effects such as chronic inflammation, granulation tissue hyperplasia and fibrosis at the implant 19 site.. In order to identify the biological responses underlying this process we developed an in vitro 20 contact-compression model that applies variable compressive stress to bronchial epithelial cells.

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1-2 h incubation. ..

    Positron Emission Tomography:

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1–2 h incubation. ..

    Article Title: Contact-compression induces inflammatory and remodeling responses in bronchial epithelial cells.
    Article Snippet: 16 Aims and objectives: Lung implantable devices, such as stents and valves are used as treatment for lung 17 cancer and COPD.. They apply continuous compressive stress to airway tissue, potentially triggering 18 adverse effects such as chronic inflammation, granulation tissue hyperplasia and fibrosis at the implant 19 site.. In order to identify the biological responses underlying this process we developed an in vitro 20 contact-compression model that applies variable compressive stress to bronchial epithelial cells.

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1-2 h incubation. ..

    Pore Size:

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1–2 h incubation. ..

    Article Title: Modulating food intake by nasal application of peptides targeting melanocortin 4 receptor and ghrelin receptor systems
    Article Snippet: .. Apparent permeability ( P app ) was measured on Calu-3 cells differentiated in translucent 12-well ThinCert inserts with a pore size of 0.4 μm (Greiner Bio-One). ..

    Article Title: Biodegradable Carbonate Nanogels Loaded with Anti MFAP-5 siRNA for Anti-stromal Therapy of Hepatocellular Carcinoma.
    Article Snippet: .. NIH/3T3 cells (10,000 cells/well) were seeded into 12-well plates, while Dt81Hepa1-6 cells (5,000 cells/insert) were seeded in cell culture inserts (ThinCert, 8 μm pore size, Greiner, Kremsmünster, Austria) and placed in the same wells for coculture. .. After 5 days of co-cultivation at 37 ◦C with 5% CO2 , NG:siMFAP5 was added at final siRNA concentrations of 25, 50, and 100 n m . For positive controls, cells were treated with anti-MFAP5 siRNA using the transfection agent Dharmafect (Dharmacon, Lafayette, USA) at 100 n m siRNA, following the manufacturer’s protocol.

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1-2 h incubation. ..

    Purification:

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1–2 h incubation. ..

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1-2 h incubation. ..

    Sterility:

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1–2 h incubation. ..

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1-2 h incubation. ..

    Incubation:

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1–2 h incubation. ..

    Article Title: Characterization of the clade 4 non-toxigenic C. difficile isolate L-NTCD03 carrying the cfr (B) gene
    Article Snippet: .. The apical side of Thincert 12 well cell culture inserts (PET, pore size 0.4 μm, Greiner Bio-one) were coated with 250 μg/well purified human collagen type IV (OptiCol collagen Type IV, dissolved in 0.25% acetic acid, Cell Guidance Systems) and washed with sterile PBS after 1-2 h incubation. ..

    other:

    Article Title: Inflammation drives TGFβ1 activation via the αvβ6 integrin-mechanotransduction pathway in human skin
    Article Snippet: Thincert chambers for RHE models , Greiner Bio-One , Cat# 665640.

    Permeability:

    Article Title: Modulating food intake by nasal application of peptides targeting melanocortin 4 receptor and ghrelin receptor systems
    Article Snippet: .. Apparent permeability ( P app ) was measured on Calu-3 cells differentiated in translucent 12-well ThinCert inserts with a pore size of 0.4 μm (Greiner Bio-One). ..

    Membrane:

    Article Title: Contact-compression induces inflammatory and remodeling responses in bronchial epithelial cells.
    Article Snippet: 16 Aims and objectives: Lung implantable devices, such as stents and valves are used as treatment for lung 17 cancer and COPD.. They apply continuous compressive stress to airway tissue, potentially triggering 18 adverse effects such as chronic inflammation, granulation tissue hyperplasia and fibrosis at the implant 19 site.. In order to identify the biological responses underlying this process we developed an in vitro 20 contact-compression model that applies variable compressive stress to bronchial epithelial cells.

    Concentration Assay:

    Article Title: Oxysterol-mediated modulation of intestinal inflammation: insights into sex differences and GPR183 signaling.
    Article Snippet: .. The cells were seeded at a concentration of 5 × 105 onto 12-well Thincerts with a 0.4 μm pore diameter (Greiner, cat# 665640), with 500 μL of medium in the apical AR TIC LE IN PR ES S 12 compartment and 1.5 mL in the basolateral compartment. ..



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    Image Search Results


    Visualization of unstained cells under microscope in green light growing on a polycarbonate ( A ) and a polyester ( B ) Transwell membrane.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: Visualization of unstained cells under microscope in green light growing on a polycarbonate ( A ) and a polyester ( B ) Transwell membrane.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques: Microscopy, Membrane

    Visualization of unstained cells growing on a polyester Transwell membrane. Left, cells attached to collagen and poly-L-lysine-coated plates; right, cells attached to collagen-, fibronectin-, and poly-L-lysine-coated plates. The same plates were observed under green light but without any fluorescence treatment.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: Visualization of unstained cells growing on a polyester Transwell membrane. Left, cells attached to collagen and poly-L-lysine-coated plates; right, cells attached to collagen-, fibronectin-, and poly-L-lysine-coated plates. The same plates were observed under green light but without any fluorescence treatment.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques: Membrane, Fluorescence

    TEER in astrocyte and pericyte co-cultures seeded in a polyester Transwell insert. Data represent values from triplicate wells on each day for 6 days after seeding. The star indicates the cell density that was considered optimal and carried forward for further experiments. TEER, transepithelial electrical resistance.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: TEER in astrocyte and pericyte co-cultures seeded in a polyester Transwell insert. Data represent values from triplicate wells on each day for 6 days after seeding. The star indicates the cell density that was considered optimal and carried forward for further experiments. TEER, transepithelial electrical resistance.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques:

    TEER in four-cell model containing hBMECs, astrocytes, pericytes, and neurons in a polyester Transwell insert either according to the standard setup or with changes to various conditions: ± Zn; ± serum; ± neurons; or hBMECs only. At each point, triplicates are used for rigor and TEER was measured three times. On day 11 after cell seeding, cells on the Transwell insert membrane were fixed and preserved for confocal microscopy study. TEER, trans-epithelial electrical resistance; Zn, zinc; hBMECs, human brain microvascular endothelial cells.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: TEER in four-cell model containing hBMECs, astrocytes, pericytes, and neurons in a polyester Transwell insert either according to the standard setup or with changes to various conditions: ± Zn; ± serum; ± neurons; or hBMECs only. At each point, triplicates are used for rigor and TEER was measured three times. On day 11 after cell seeding, cells on the Transwell insert membrane were fixed and preserved for confocal microscopy study. TEER, trans-epithelial electrical resistance; Zn, zinc; hBMECs, human brain microvascular endothelial cells.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques: Membrane, Confocal Microscopy

    Expression of tight junctions (TJs), ZO-1, and claudin 5 on human brain cells, layered on polyester Transwell insert membrane. Staining was performed for ZO-1 (red), claudin 5 (green), and the nucleus (DAPI; blue). TJs were observed on astrocytes and pericytes growing in medium alone ( A ) or with added zinc sulfate ( B ) and on hBMECs in medium alone ( C ) or with zinc sulfate ( D ). Scale bar inside images represents 50 μm. TJ, tight junction; Z0–1, zona occludens; hBMEC, human brain microvascular endothelial cells; DAPI, 4′,6-diamidino-2-phenylindole.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: Expression of tight junctions (TJs), ZO-1, and claudin 5 on human brain cells, layered on polyester Transwell insert membrane. Staining was performed for ZO-1 (red), claudin 5 (green), and the nucleus (DAPI; blue). TJs were observed on astrocytes and pericytes growing in medium alone ( A ) or with added zinc sulfate ( B ) and on hBMECs in medium alone ( C ) or with zinc sulfate ( D ). Scale bar inside images represents 50 μm. TJ, tight junction; Z0–1, zona occludens; hBMEC, human brain microvascular endothelial cells; DAPI, 4′,6-diamidino-2-phenylindole.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques: Expressing, Membrane, Staining

    Expression of ZO-1 (red) and claudin 5 (green) in monoculture of hBMECs on a Transwell membrane ( A ) and determination of claudin 5 on neuron cell from the non-contact four-cell model with neuronal cell marker in red ( B ) and when the medium for the BBB model was supplemented with Zn ( C ). Scale bars: 20 μm in ( A ) and ( B ), 50 μm in ( C ). Zn, zinc; Z0–1, zona occludens; hBMECs, human brain microvascular endothelial cells; DAPI, 4′,6-diamidino-2-phenylindole.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: Expression of ZO-1 (red) and claudin 5 (green) in monoculture of hBMECs on a Transwell membrane ( A ) and determination of claudin 5 on neuron cell from the non-contact four-cell model with neuronal cell marker in red ( B ) and when the medium for the BBB model was supplemented with Zn ( C ). Scale bars: 20 μm in ( A ) and ( B ), 50 μm in ( C ). Zn, zinc; Z0–1, zona occludens; hBMECs, human brain microvascular endothelial cells; DAPI, 4′,6-diamidino-2-phenylindole.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques: Expressing, Membrane, Marker

    Cell identification by detecting s100β on astrocytes and CD146 on pericytes and hBMECs as respective cell markers. Astrocytes, pericytes, and hBMECs on a Transwell membrane, immunostained with a cocktail of s100β antibody (red), an antibody against CD146 (green); the nucleus was stained with DAPI (blue). The basolateral surface of the Transwell membrane shows astrocyte and pericyte ( A ), and the apical part of the same Transwell membrane shows hBMECs only ( B ). Scale bars: 50 μm. Abbreviations: hBMECs, human brain microvascular endothelial cells; DAPI, 4′,6-diamidino-2-phenylindole.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: Cell identification by detecting s100β on astrocytes and CD146 on pericytes and hBMECs as respective cell markers. Astrocytes, pericytes, and hBMECs on a Transwell membrane, immunostained with a cocktail of s100β antibody (red), an antibody against CD146 (green); the nucleus was stained with DAPI (blue). The basolateral surface of the Transwell membrane shows astrocyte and pericyte ( A ), and the apical part of the same Transwell membrane shows hBMECs only ( B ). Scale bars: 50 μm. Abbreviations: hBMECs, human brain microvascular endothelial cells; DAPI, 4′,6-diamidino-2-phenylindole.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques: Membrane, Staining

    Average fluorescence intensity analysis of claudin 5 and drug penetration for different sets of BBB representation. ( A ) Cell surface pixel intensity for claudin 5 measured using Fiji ImageJ-win64 software for image analysis. ( B ) Evaluation of tight junction integrity and the penetration of HIV-ART drug DTG in the four-cell mode. The color-coded bar pairs show drug distribution in the apical and bottom layers of the indicated in vitro models and control (membrane only) from a Transwell membrane plate. Medium from the apical and basal layer of the Transwell membrane was collected after 48 hr of drug treatment. Data represent the mean concentrations from three replicates. MFI, mean fluorescence intensity; AP, astrocytes and pericytes; DTG, dolutegravir; hBMECs, human brain microvascular endothelial cells; Zn, zinc.

    Journal: Current protocols

    Article Title: Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells

    doi: 10.1002/cpz1.1067

    Figure Lengend Snippet: Average fluorescence intensity analysis of claudin 5 and drug penetration for different sets of BBB representation. ( A ) Cell surface pixel intensity for claudin 5 measured using Fiji ImageJ-win64 software for image analysis. ( B ) Evaluation of tight junction integrity and the penetration of HIV-ART drug DTG in the four-cell mode. The color-coded bar pairs show drug distribution in the apical and bottom layers of the indicated in vitro models and control (membrane only) from a Transwell membrane plate. Medium from the apical and basal layer of the Transwell membrane was collected after 48 hr of drug treatment. Data represent the mean concentrations from three replicates. MFI, mean fluorescence intensity; AP, astrocytes and pericytes; DTG, dolutegravir; hBMECs, human brain microvascular endothelial cells; Zn, zinc.

    Article Snippet: 70% (v/v) ethanol Endothelial CM (see recipe) Transwell inserts bearing BBB models (Basic Protocol ) Paraformaldehyde (Thermo Scientific, cat. no. J19943-K2) Dulbecco’s phosphate-buffered saline (DPBS; Gibco, cat. no. 14190144) Goat serum (Abcam, cat. no. ab7481) Tween-20 (Sigma, cat. no. P2287–100ML) Bovine serum albumin (BSA) Wash buffer (see recipe) Appropriately conjugated (labeled) antibodies for claudin 5 (Invitrogen, cat. no. 362588) and ZO-1 (Invitrogen, cat. no. MA3–39100 A647) Appropriately conjugated (labeled) antibodies for cell markers: anti-S1OOβ for astrocytes (Abcam, cat. no. ab196175), anti-CD146 (Abcam, cat. no. ab196448 for pericytes and hBMECs), and anti-NeuN antibody for neuron (Abcam, ab190565) DAPI-(4′,6-diamidino-2-phenylindole) mounting dye (ProLong Diamond Antifade Mounting medium with DAPI, Invitrogen cat. no. P3692) Dolutegravir (DTG; Adooq Bioscience) DTG solution without DTG (see recipe) EVOM2 for TEER measurement (World Precision Instruments [WPI]) Scalpel Scissors Beakers Glass slides Glass coverslips Forceps Measuring cylinders Magnetic stirrer Magnetic beads Vortex Confocal microscope (Zeiss LSM800 microscope) Fiji ImageJ software Liquid chromatography-tandem mass spectrometer (LC/MS/MS)

    Techniques: Fluorescence, Software, In Vitro, Control, Membrane