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q plex human custom cytokine plex kit  (Quansys Biosciences)


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    Quansys Biosciences q plex human custom cytokine plex kit
    Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as <LLOQ. (I) Quantification of collagen type II fragments released into the culture medium at day 21, measured by ELISA ( n = 3 independent samples per group). All fluorescence images include a 100 µm scale bar, and white dashed lines indicate microchannel boundaries. Statistical analysis was performed using the Kruskal–Wallis test with Dunn's post hoc comparison (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant). Conditions include control (no <t>cytokine)</t> and IL‑1β stimulation (1 and 10 ng/ml). Note: Panels are adapted from da Silva et al. , licensed under CC BY 3.0.
    Q Plex Human Custom Cytokine Plex Kit, supplied by Quansys Biosciences, used in various techniques. Bioz Stars score: 99/100, based on 406 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+array/Q-Plex+Custom+Array/pmc13690009-99-130-137
    Average 99 stars, based on 406 article reviews
    q plex human custom cytokine plex kit - by Bioz Stars, 2026-11
    99/100 stars

    Images

    1) Product Images from "A Gravity‐driven Microfluidic Platform for High‐throughput Culture and Inflammatory Stimulation of Human Articular Chondrocytes"

    Article Title: A Gravity‐driven Microfluidic Platform for High‐throughput Culture and Inflammatory Stimulation of Human Articular Chondrocytes

    Journal: Current Protocols

    doi: 10.1002/cpz1.70489

    Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as <LLOQ. (I) Quantification of collagen type II fragments released into the culture medium at day 21, measured by ELISA ( n = 3 independent samples per group). All fluorescence images include a 100 µm scale bar, and white dashed lines indicate microchannel boundaries. Statistical analysis was performed using the Kruskal–Wallis test with Dunn's post hoc comparison (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant). Conditions include control (no cytokine) and IL‑1β stimulation (1 and 10 ng/ml). Note: Panels are adapted from da Silva et al. , licensed under CC BY 3.0.
    Figure Legend Snippet: Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as

    Techniques Used: Morphology, Expressing, Activity Assay, Fluorescence, Cell Culture, Shear, Staining, Control, Immunofluorescence, Marker, Prestoblue Assay, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Analysis, Comparison

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: AGER-1 Long Non-Coding RNA Levels Correlate with the Expression of the Advanced Glycosylation End-Product Receptor, a Regulator of the Inflammatory Response in Visceral Adipose Tissue of Women with Obesity and Type 2 Diabetes Mellitus.
    Article Snippet: .. An ELISA-based chemiluminescent Q-plex Custom array (Quansys Bioscience, West Logan, UT, USA) was used to measure interleukin (IL) 1β, 6, 8, and resistin concentrations in adipose tissue protein extracts. .. Luminescence was assessed using a Molecular Imager Versa DocTM MP 5000 system (Bio-Rad, Hercules, CA, USA), according to the manufacturer’s guidelines.

    Article Title: Interleukin-23 levels in umbilical cord blood are associated with neurodevelopmental trajectories in infancy
    Article Snippet: .. Serum concentrations of interleulin (IL)-1α, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-23, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, TNF-β, eotaxin, C-C motif chemokine ligand (CCL)-2, CCL-3, growth-regulated oncogene (GRO)α, I-309, interferon-inducible protein (IP)-10, regulated on activation, normal T cell expressed and secreted (RANTES), thymus and activation-regulated chemokine (TARC), angiopoietin (Ang)-2, fibroblast growth factor (FGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), tissue inhibitor of metalloproteinase (TIMP)-1, TIMP2, and vascular endothelial growth factor (VEGF) were determined using a Q-Plex Human Customs Array multiplexed ELISA (Quansys Biosciences, UT.). ..

    Article Title: The Role of microRNA in the Regulation of Cortisol Metabolism in the Adipose Tissue in the Course of Obesity
    Article Snippet: .. Interleukin (IL) 1β, 6, 8, 15, adiponectin, and resistin concentrations in adipose tissue protein extracts were measured using an ELISA-based chemiluminescent Q-plex Custom array (Quansys Bioscience, West Logan, UT, USA). .. A Molecular Imager Versa DocTM MP 5000 system (Bio-Rad, Hercules, CA, USA) was used to assess luminescence according to the manufacturer’s guidelines.

    Activation Assay:

    Article Title: Interleukin-23 levels in umbilical cord blood are associated with neurodevelopmental trajectories in infancy
    Article Snippet: .. Serum concentrations of interleulin (IL)-1α, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-23, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, TNF-β, eotaxin, C-C motif chemokine ligand (CCL)-2, CCL-3, growth-regulated oncogene (GRO)α, I-309, interferon-inducible protein (IP)-10, regulated on activation, normal T cell expressed and secreted (RANTES), thymus and activation-regulated chemokine (TARC), angiopoietin (Ang)-2, fibroblast growth factor (FGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), tissue inhibitor of metalloproteinase (TIMP)-1, TIMP2, and vascular endothelial growth factor (VEGF) were determined using a Q-Plex Human Customs Array multiplexed ELISA (Quansys Biosciences, UT.). ..

    Sterility:

    Article Title: A Gravity‐driven Microfluidic Platform for High‐throughput Culture and Inflammatory Stimulation of Human Articular Chondrocytes
    Article Snippet: .. Chondrocyte growth medium (HC Growth Medium, Cell Applications, cat. no. 411‐500) DPBS (sterile, Gibco, cat. no. 14190144) Recombinant IL‐1β (R&D Systems, cat. no. 201‐LB‐025) (separately at −20°C) Live/Dead Viability/Cytotoxicity Kit containing calcein‐AM and ethidium homodimer 1 (EthD‐1) (Invitrogen, cat. no. LS224) PrestoBlue Cell Viability Reagent (Invitrogen, cat. no. A13261) 4% paraformaldehyde (PFA) in DPBS (Thermo Scientific, cat. no. J61899.AP) Normal donkey serum Bovine serum albumin (BSA) Triton X‐100 Tetramethylrhodamine (TRITC)‐phalloidin staining (EMD Millipore/Merck, cat. no. 90228) Mouse anti‐Collagen II antibody (NB600‐844, Novus Biologicals) Goat anti‐Aggrecan antibody (AF1220, R&D Systems) Rabbit anti‐Mouse IgG (H+L), FITC‐conjugated secondary antibody (NB7543, Novus Biologicals) Donkey anti‐Goat IgG (H+L), Northern Lights 557 (NL557)‐conjugated secondary antibody (NL001, R&D Systems) 4′,6‐diamidino‐2‐phenylindole (DAPI) Solution (Novus Biologicals, Bio‐Techne, cat. no. NBP2‐31156) Q‐Plex Human MMP 6 Plex kit (340949HU, Quansys Biosciences) Q‐Plex Human Custom Cytokine Plex kit (107749GR, Quansys Biosciences) Human collagen type II alpha 1 ELISA kit (BSKH63293, Bioss Antibodies) Pre‐established chondrocyte cultures generated using Basic Protocol Rocker platform (AKITA rocker, FinnAdvance or equivalent) CO incubator maintained at 37°C and 5% CO Class II biosafety cabinet Wide‐field fluorescence microscope (e.g., Leica DM4B or equivalent) equipped with blue/green and red/green excitation/emission filter sets. .. Wide‐field fluorescence microscope equipped for z‐stack imaging (e.g., Leica DM6B Thunder Microscope or equivalent) with UV/blue, blue/green, and red/green excitation/emission filter sets.



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    Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as <LLOQ. (I) Quantification of collagen type II fragments released into the culture medium at day 21, measured by ELISA ( n = 3 independent samples per group). All fluorescence images include a 100 µm scale bar, and white dashed lines indicate microchannel boundaries. Statistical analysis was performed using the Kruskal–Wallis test with Dunn's post hoc comparison (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant). Conditions include control (no <t>cytokine)</t> and IL‑1β stimulation (1 and 10 ng/ml). Note: Panels are adapted from da Silva et al. , licensed under CC BY 3.0.
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    Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as <LLOQ. (I) Quantification of collagen type II fragments released into the culture medium at day 21, measured by ELISA ( n = 3 independent samples per group). All fluorescence images include a 100 µm scale bar, and white dashed lines indicate microchannel boundaries. Statistical analysis was performed using the Kruskal–Wallis test with Dunn's post hoc comparison (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant). Conditions include control (no <t>cytokine)</t> and IL‑1β stimulation (1 and 10 ng/ml). Note: Panels are adapted from da Silva et al. , licensed under CC BY 3.0.
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    Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as <LLOQ. (I) Quantification of collagen type II fragments released into the culture medium at day 21, measured by ELISA ( n = 3 independent samples per group). All fluorescence images include a 100 µm scale bar, and white dashed lines indicate microchannel boundaries. Statistical analysis was performed using the Kruskal–Wallis test with Dunn's post hoc comparison (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant). Conditions include control (no <t>cytokine)</t> and IL‑1β stimulation (1 and 10 ng/ml). Note: Panels are adapted from da Silva et al. , licensed under CC BY 3.0.
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    Image Search Results


    Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as <LLOQ. (I) Quantification of collagen type II fragments released into the culture medium at day 21, measured by ELISA ( n = 3 independent samples per group). All fluorescence images include a 100 µm scale bar, and white dashed lines indicate microchannel boundaries. Statistical analysis was performed using the Kruskal–Wallis test with Dunn's post hoc comparison (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; n.s., not significant). Conditions include control (no cytokine) and IL‑1β stimulation (1 and 10 ng/ml). Note: Panels are adapted from da Silva et al. , licensed under CC BY 3.0.

    Journal: Current Protocols

    Article Title: A Gravity‐driven Microfluidic Platform for High‐throughput Culture and Inflammatory Stimulation of Human Articular Chondrocytes

    doi: 10.1002/cpz1.70489

    Figure Lengend Snippet: Expected cellular responses of human articular chondrocytes under dynamic culture and IL‑1β‐induced inflammatory conditions. Representative results illustrate chondrocyte morphology, viability, extracellular matrix (ECM) expression, metabolic activity, and inflammatory responses under gravity‐driven flow conditions. (A) Representative z‐projection fluorescence images of chondrocytes cultured for 21 days in microchannels coated with a fibronectin–collagen type IV mixture. Cells form aggregates and microtissue‐like structures across regions I–III corresponding to increasing fluid shear stress levels. Cells are stained for F‐actin (red) and nuclei (DAPI, blue). (B) Representative Live/Dead fluorescence images showing time‐dependent viability and proliferation of chondrocytes cultured under dynamic conditions on days 3, 7, 14, and 21 (control), compared with IL‑1β‐stimulated cultures on day 21 (1 and 10 ng/ml, applied from day 15 to day 21). Control cultures show progressive formation of compact microtissue‐like aggregates (white arrows), whereas IL‑1β stimulation results in disrupted aggregates (yellow arrows). Green: calcein‐AM (live cells); red: ethidium homodimer‐1 (dead cells). (C) Representative z‐projection immunofluorescence images of ECM marker expression in region I after 21 days of culture, comparing control and IL‑1β‐stimulated samples. Images show merged fluorescence channels for collagen type II (green), aggrecan (red), and nuclei (DAPI, blue). (D–E) Quantification of corrected total cell fluorescence (CTCF) for collagen type II (D) and aggrecan (E) across regions I–III for control and IL‑1β‐stimulated cultures at day 21 (n = 44 cells per group). (F) Cell metabolic activity measured using a PrestoBlue assay for control and IL‑1β‐stimulated samples at day 21 ( n = 5 independent samples per group). (G–H) Quantification of inflammatory mediators in culture supernatant collected at day 21 using multiplex ELISA, including matrix metalloproteinases (MMP‐1, MMP‐2, MMP‐3, MMP‐7, MMP‐9, MMP‐13) and cytokines (IL‑1β, IL‑6, IL‑8) ( n = 3 independent samples per group). Values below the lower limit of quantification are indicated as

    Article Snippet: Chondrocyte growth medium (HC Growth Medium, Cell Applications, cat. no. 411‐500) DPBS (sterile, Gibco, cat. no. 14190144) Recombinant IL‐1β (R&D Systems, cat. no. 201‐LB‐025) (separately at −20°C) Live/Dead Viability/Cytotoxicity Kit containing calcein‐AM and ethidium homodimer 1 (EthD‐1) (Invitrogen, cat. no. LS224) PrestoBlue Cell Viability Reagent (Invitrogen, cat. no. A13261) 4% paraformaldehyde (PFA) in DPBS (Thermo Scientific, cat. no. J61899.AP) Normal donkey serum Bovine serum albumin (BSA) Triton X‐100 Tetramethylrhodamine (TRITC)‐phalloidin staining (EMD Millipore/Merck, cat. no. 90228) Mouse anti‐Collagen II antibody (NB600‐844, Novus Biologicals) Goat anti‐Aggrecan antibody (AF1220, R&D Systems) Rabbit anti‐Mouse IgG (H+L), FITC‐conjugated secondary antibody (NB7543, Novus Biologicals) Donkey anti‐Goat IgG (H+L), Northern Lights 557 (NL557)‐conjugated secondary antibody (NL001, R&D Systems) 4′,6‐diamidino‐2‐phenylindole (DAPI) Solution (Novus Biologicals, Bio‐Techne, cat. no. NBP2‐31156) Q‐Plex Human MMP 6 Plex kit (340949HU, Quansys Biosciences) Q‐Plex Human Custom Cytokine Plex kit (107749GR, Quansys Biosciences) Human collagen type II alpha 1 ELISA kit (BSKH63293, Bioss Antibodies) Pre‐established chondrocyte cultures generated using Basic Protocol Rocker platform (AKITA rocker, FinnAdvance or equivalent) CO incubator maintained at 37°C and 5% CO Class II biosafety cabinet Wide‐field fluorescence microscope (e.g., Leica DM4B or equivalent) equipped with blue/green and red/green excitation/emission filter sets.

    Techniques: Morphology, Expressing, Activity Assay, Fluorescence, Cell Culture, Shear, Staining, Control, Immunofluorescence, Marker, Prestoblue Assay, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Analysis, Comparison