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well boyden chamber model  (Guangzhou JET Bio-Filtration)


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    Structured Review

    Guangzhou JET Bio-Filtration well boyden chamber model
    Well Boyden Chamber Model, supplied by Guangzhou JET Bio-Filtration, used in various techniques. Bioz Stars score: 99/100, based on 103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/insertion+model/Tissue+Culture+Plate+Inserts/pm39910045-279-14-25
    Average 99 stars, based on 103 article reviews
    well boyden chamber model - by Bioz Stars, 2026-11
    99/100 stars

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    Related Articles

    other:

    Article Title: Tissue-specific inflammation induces cell state plasticity with oncogenic addiction in mucosal melanoma.
    Article Snippet: Luminescence was measured using a Spark microplate reader (Tecan), and cell viability was calculated using the following formula The chemicals and recombinant proteins used in this study are listed in Table 2.

    Migration:

    Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation
    Article Snippet: .. Cell migration assays were performed transwell inserts (TCS003024, Jet Biofil, Guangzhou, China). ..

    Article Title: Environmental benzophenone exposure promotes breast cancer cell metastasis via the Wnt/β-catenin pathway.
    Article Snippet: Humans, in particular women, are widely exposed to benzophenones, a major supplement in personal care products.. However, little is known about the impact of chronic benzophenone exposure on breast cancer progression.. In this study, we applied a long-term cell culture model by treating the triple negative breast cancer MDA-MB-231 cells to environmental dosage of 4-hydroxybenzophenone (4HBP), the main metabolite of benzophenones.

    Pore Size:

    Article Title: Environmental benzophenone exposure promotes breast cancer cell metastasis via the Wnt/β-catenin pathway.
    Article Snippet: Humans, in particular women, are widely exposed to benzophenones, a major supplement in personal care products.. However, little is known about the impact of chronic benzophenone exposure on breast cancer progression.. In this study, we applied a long-term cell culture model by treating the triple negative breast cancer MDA-MB-231 cells to environmental dosage of 4-hydroxybenzophenone (4HBP), the main metabolite of benzophenones.

    Article Title: Tissue-specific inflammation induces cell state plasticity with oncogenic addiction in mucosal melanoma
    Article Snippet: M1 polarization was achieved by supplementation with IFN-γ (50 ng/ml; PeproTech, 300-02) and LPS (20 ng/ml; Sigma-Aldrich, L2630) for another 24 hours. .. In the coculture experiment, monocytes were differentiated into M1 macrophages in a 24-well plate, whereas MMs were plated in the 24 Transwell inserts (membrane pore size of 0.4 μm; Jet Biofil, TCS021024). .. RMPI 1640 supplemented with macrophage colony-stimulating factor (M-CSF) (100 ng/ml; TargetMoI, TMPY-00127), 10% heat-inactivated FBS, 1% glutamine, and 1% penicillin-streptomycin was added to the plates and incubated at 5% CO 2 at 37°C for 7 days to differentiate monocytes into nonpolarized macrophages.

    Article Title: In vitro iron bioaccessibility and intestinal transport of the edible red alga Bangia atropurpurea: Molecular-weight distribution and cellular metabolic response
    Article Snippet: The nutritional value of iron-rich algae depends on iron release during digestion and subsequent intestinal availability.. This study evaluated iron from the edible red alga Bangia atropurpurea using complementary physicochemical and intestinal in vitro models.. The analyzed commercial material contained 486 ± 77 μg Fe/g dry weight.

    Article Title: Tissue-specific inflammation induces cell state plasticity with oncogenic addiction in mucosal melanoma
    Article Snippet: Macrophages were further polarized in M1 macrophages by incubation with interferon-γ (IFN-γ) (20 ng/ml; PeproTech, 300-02) and LPS (10 ng/ml; Sigma-Aldrich, L2630) for another 24 hours. .. In the coculture experiment, THP-1 monocytes were differentiated into M1 macrophages in a 24-well plate, whereas MMs were plated in the 24 Transwell inserts (membrane pore size of 0.4 μm; Jet Biofil, TCS021024). .. THP-1 monocytes were differentiated into M0 macrophages by 48 hours of incubation with phorbol 12-myristate 13-acetate (PMA; 100 ng/ml; Yeasen, 50601ES03).

    Membrane:

    Article Title: Tissue-specific inflammation induces cell state plasticity with oncogenic addiction in mucosal melanoma
    Article Snippet: M1 polarization was achieved by supplementation with IFN-γ (50 ng/ml; PeproTech, 300-02) and LPS (20 ng/ml; Sigma-Aldrich, L2630) for another 24 hours. .. In the coculture experiment, monocytes were differentiated into M1 macrophages in a 24-well plate, whereas MMs were plated in the 24 Transwell inserts (membrane pore size of 0.4 μm; Jet Biofil, TCS021024). .. RMPI 1640 supplemented with macrophage colony-stimulating factor (M-CSF) (100 ng/ml; TargetMoI, TMPY-00127), 10% heat-inactivated FBS, 1% glutamine, and 1% penicillin-streptomycin was added to the plates and incubated at 5% CO 2 at 37°C for 7 days to differentiate monocytes into nonpolarized macrophages.

    Article Title: Tissue-specific inflammation induces cell state plasticity with oncogenic addiction in mucosal melanoma
    Article Snippet: Macrophages were further polarized in M1 macrophages by incubation with interferon-γ (IFN-γ) (20 ng/ml; PeproTech, 300-02) and LPS (10 ng/ml; Sigma-Aldrich, L2630) for another 24 hours. .. In the coculture experiment, THP-1 monocytes were differentiated into M1 macrophages in a 24-well plate, whereas MMs were plated in the 24 Transwell inserts (membrane pore size of 0.4 μm; Jet Biofil, TCS021024). .. THP-1 monocytes were differentiated into M0 macrophages by 48 hours of incubation with phorbol 12-myristate 13-acetate (PMA; 100 ng/ml; Yeasen, 50601ES03).

    Tissue:

    Article Title: Carrier-free ternary self-assembled nanoparticle hydrogel dual-targeting AGEs and STING pathways to accelerate diabetic wound healing
    Article Snippet: Excessive accumulation of advanced glycation end-products (AGEs) and aberrant activation of the stimulator of interferon genes (STING) pathway are two major pathological drivers of angiogenic dysfunction and persistent inflammation in diabetic wounds (DW), which account for the refractory nature of DW.. Herein, we develop a multifunctional hydrogel (CE-Gel) fabricated from chitooligosaccharide (COS) and silk fibroin (SF), loaded with carrier-free ternary self-assembled nanoparticles (ESF NPs) composed of epigallocatechin gallate (EGCG), the inhibitor (FPS-ZM1) of the receptor for AGEs (RAGE) and the STING inhibitor (SN001).. CE-Gel promotes DW healing via dual-targeting both on AGEs and STING pathways.

    Cell Culture:

    Article Title: In vitro iron bioaccessibility and intestinal transport of the edible red alga Bangia atropurpurea: Molecular-weight distribution and cellular metabolic response
    Article Snippet: The nutritional value of iron-rich algae depends on iron release during digestion and subsequent intestinal availability.. This study evaluated iron from the edible red alga Bangia atropurpurea using complementary physicochemical and intestinal in vitro models.. The analyzed commercial material contained 486 ± 77 μg Fe/g dry weight.



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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