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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

    Multiplex Assay:

    Article Title: Thiophene-Benzimidazole Conjugates Attenuate IL-33-induced Inflammation in Human Mast Cell Line and in ex vivo Human Skin Organ Cultures.
    Article Snippet: IL-6 and IL-8 kits were from Bioligand cat (BLG-430515 and BLG-431504, respectively). .. Quansys biosciences supplied the multiplex array (cat: 112433HU). .. Total RNA Mini Plus Concentrator kit (cat#: 036-100-M2 A&A Biotechnology), qPCR Kits: GoTaq qPCR and RT-qPCR Systems (cat#: A6020) were from Promega.

    other:

    Article Title: Iron status at early pregnancy is associated with infectious respiratory and gastric illness in women receiving routine iron supplementation: the NuPED prospective cohort.
    Article Snippet: Ferritin, sTfR and inflammatory markers (α1-acid glycoprotein [AGP] and C-reactive protein [CRP]) concentrations were measured with the Q-Plex Human Micronutrient Array (7-plex, Quansys Bioscience, Logan, UT, USA) [18].

    Concentration Assay:

    Article Title: Performance and usability evaluation of three LDH-based malaria rapid diagnostic tests in Kédougou, Senegal.
    Article Snippet: All assays were performed using Agilent Mx3005pro thermocyclers (Agilent Technologies, Santa Clara, CA, USA). .. Reference antigen concentration determination Antigen concentration was determined with a quantitative antigen assay: Q-PlexTM Human Malaria Array (5-Plex) (Quansys Biosciences, Logan UT, USA). ..

    Article Title: Vitamin A intake and status, and associations with iron status, anaemia, and birth outcomes of pregnant women in the Free State Province, South Africa: The NuEMI study.
    Article Snippet: .. RBP4, markers of niversity Press iron status, i.e., ferritin and soluble transferrin receptor (sTfR) concentration, and markers of inflammation, i.e., C-reactive protein (CRP) and alpha-1-acid glycoprotein (AGP) were measured in 50 μL of plasma using the Q-PlexTM Human Micronutrient Array (7-plex) (Quansys Bioscience, UT, USA) (31) at the micronutrient laboratory of CEN. ..

    Article Title: Performance and usability evaluation of three LDH-based malaria rapid diagnostic tests in Kédougou, Senegal
    Article Snippet: All assays were performed using Agilent Mx3005pro thermocyclers (Agilent Technologies, Santa Clara, CA, USA). .. Antigen concentration was determined with a quantitative antigen assay: Q-PlexTM Human Malaria Array (5-Plex) (Quansys Biosciences, Logan UT, USA). ..

    Antigen Assay:

    Article Title: Performance and usability evaluation of three LDH-based malaria rapid diagnostic tests in Kédougou, Senegal.
    Article Snippet: All assays were performed using Agilent Mx3005pro thermocyclers (Agilent Technologies, Santa Clara, CA, USA). .. Reference antigen concentration determination Antigen concentration was determined with a quantitative antigen assay: Q-PlexTM Human Malaria Array (5-Plex) (Quansys Biosciences, Logan UT, USA). ..

    Article Title: Performance and usability evaluation of three LDH-based malaria rapid diagnostic tests in Kédougou, Senegal
    Article Snippet: All assays were performed using Agilent Mx3005pro thermocyclers (Agilent Technologies, Santa Clara, CA, USA). .. Antigen concentration was determined with a quantitative antigen assay: Q-PlexTM Human Malaria Array (5-Plex) (Quansys Biosciences, Logan UT, USA). ..

    Clinical Proteomics:

    Article Title: Vitamin A intake and status, and associations with iron status, anaemia, and birth outcomes of pregnant women in the Free State Province, South Africa: The NuEMI study.
    Article Snippet: .. RBP4, markers of niversity Press iron status, i.e., ferritin and soluble transferrin receptor (sTfR) concentration, and markers of inflammation, i.e., C-reactive protein (CRP) and alpha-1-acid glycoprotein (AGP) were measured in 50 μL of plasma using the Q-PlexTM Human Micronutrient Array (7-plex) (Quansys Bioscience, UT, USA) (31) at the micronutrient laboratory of CEN. ..

    Article Title: Culturally acceptable fermented grain may improve gut health in South African postpartum mothers in a randomised trial.
    Article Snippet: .. Maternal plasma collected at weeks 4 and 10 was evaluated for soluble transferrin receptor (sTfR, Fe sufficiency), retinol binding protein 4 (RBP4, indicator of vitamin A and protein status), thyroglobulin (Tg, iodine status), C-reactive protein (CRP) and α-1-acid glycoprotein (systemic inflammation) using the Q-Plex Human Micronutrient array (Quansys Biosciences). .. 25-Hydroxy vitamin D (25(OH)VitD3), vitamin B12, ferritin and Fe levels were measured in maternal serum collected at weeks 4 and 10 by the local National Health Laboratory Service Pathology lab in Cape Town, South Africa.

    Binding Assay:

    Article Title: Culturally acceptable fermented grain may improve gut health in South African postpartum mothers in a randomised trial.
    Article Snippet: .. Maternal plasma collected at weeks 4 and 10 was evaluated for soluble transferrin receptor (sTfR, Fe sufficiency), retinol binding protein 4 (RBP4, indicator of vitamin A and protein status), thyroglobulin (Tg, iodine status), C-reactive protein (CRP) and α-1-acid glycoprotein (systemic inflammation) using the Q-Plex Human Micronutrient array (Quansys Biosciences). .. 25-Hydroxy vitamin D (25(OH)VitD3), vitamin B12, ferritin and Fe levels were measured in maternal serum collected at weeks 4 and 10 by the local National Health Laboratory Service Pathology lab in Cape Town, South Africa.

    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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    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