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diseased ipf lung fibroblasts  (BioIVT Inc)

 
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    BioIVT Inc diseased ipf lung fibroblasts
    DNA damage is a hallmark of commercial primary <t>IPF</t> <t>fibroblasts</t> in vitro , but not Cdkn2a /p16 ink4a expression or a fibrotic secretome . ( A ) Primary NHLF and IPF cells ( n = 3 different donors, each with 2 technical replicates) were cultured at the same density overnight on standard tissue culture plates in low-serum growth medium and were subsequently fixed. Immunocytochemistry was performed to fluorescently label nuclei (blue) and p21 Waf1/Cip1 (orange). Images were acquired with a 40X water objective using an Operetta High Content Screening instrument and intensity of nuclear p21 Waf1/Cip1 was quantified and calculated as a positive percentage of each population. ( B ) NHLF and IPF cells were treated and imaged as in ( A ), and immunocytochemistry was performed to fluorescently label nuclei (blue) and DNA damage via nuclear γH2A.X (Ser139) foci (orange). The number of nuclear foci per cell was quantified and cells with one or more were reported as positive. ( C ) Following overnight culture in standard tissue culture plates, NHLF and IPF cells were lysed, and qPCR was performed. Expression of senescence and fibrosis-related matrix and secreted factor genes was assessed, and data were normalized to β2m housekeeper expression using the 2 −ΔΔCt method versus NHLF cells. ( D ) Following overnight culture as in ( C ), cell culture supernatants were collected and assayed for determination of the concentration of common fibrosis-related secreted proteins by MSD kits. Statistical analysis was performed using an unpaired t -test ( A , B ) or a two-way ANOVA with a Bonferroni post-test ( C , D ) in GraphPad Prism: * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent standard error of the mean (SEM).
    Diseased Ipf Lung Fibroblasts, supplied by BioIVT Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ipf+fibroblasts/diseased+ipf+lung+fibroblasts/pmc11272128-159-13-20
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    diseased ipf lung fibroblasts - by Bioz Stars, 2026-10
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    1) Product Images from "Modulating in vitro lung fibroblast activation via senolysis of senescent human alveolar epithelial cells"

    Article Title: Modulating in vitro lung fibroblast activation via senolysis of senescent human alveolar epithelial cells

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.205994

    DNA damage is a hallmark of commercial primary IPF fibroblasts in vitro , but not Cdkn2a /p16 ink4a expression or a fibrotic secretome . ( A ) Primary NHLF and IPF cells ( n = 3 different donors, each with 2 technical replicates) were cultured at the same density overnight on standard tissue culture plates in low-serum growth medium and were subsequently fixed. Immunocytochemistry was performed to fluorescently label nuclei (blue) and p21 Waf1/Cip1 (orange). Images were acquired with a 40X water objective using an Operetta High Content Screening instrument and intensity of nuclear p21 Waf1/Cip1 was quantified and calculated as a positive percentage of each population. ( B ) NHLF and IPF cells were treated and imaged as in ( A ), and immunocytochemistry was performed to fluorescently label nuclei (blue) and DNA damage via nuclear γH2A.X (Ser139) foci (orange). The number of nuclear foci per cell was quantified and cells with one or more were reported as positive. ( C ) Following overnight culture in standard tissue culture plates, NHLF and IPF cells were lysed, and qPCR was performed. Expression of senescence and fibrosis-related matrix and secreted factor genes was assessed, and data were normalized to β2m housekeeper expression using the 2 −ΔΔCt method versus NHLF cells. ( D ) Following overnight culture as in ( C ), cell culture supernatants were collected and assayed for determination of the concentration of common fibrosis-related secreted proteins by MSD kits. Statistical analysis was performed using an unpaired t -test ( A , B ) or a two-way ANOVA with a Bonferroni post-test ( C , D ) in GraphPad Prism: * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent standard error of the mean (SEM).
    Figure Legend Snippet: DNA damage is a hallmark of commercial primary IPF fibroblasts in vitro , but not Cdkn2a /p16 ink4a expression or a fibrotic secretome . ( A ) Primary NHLF and IPF cells ( n = 3 different donors, each with 2 technical replicates) were cultured at the same density overnight on standard tissue culture plates in low-serum growth medium and were subsequently fixed. Immunocytochemistry was performed to fluorescently label nuclei (blue) and p21 Waf1/Cip1 (orange). Images were acquired with a 40X water objective using an Operetta High Content Screening instrument and intensity of nuclear p21 Waf1/Cip1 was quantified and calculated as a positive percentage of each population. ( B ) NHLF and IPF cells were treated and imaged as in ( A ), and immunocytochemistry was performed to fluorescently label nuclei (blue) and DNA damage via nuclear γH2A.X (Ser139) foci (orange). The number of nuclear foci per cell was quantified and cells with one or more were reported as positive. ( C ) Following overnight culture in standard tissue culture plates, NHLF and IPF cells were lysed, and qPCR was performed. Expression of senescence and fibrosis-related matrix and secreted factor genes was assessed, and data were normalized to β2m housekeeper expression using the 2 −ΔΔCt method versus NHLF cells. ( D ) Following overnight culture as in ( C ), cell culture supernatants were collected and assayed for determination of the concentration of common fibrosis-related secreted proteins by MSD kits. Statistical analysis was performed using an unpaired t -test ( A , B ) or a two-way ANOVA with a Bonferroni post-test ( C , D ) in GraphPad Prism: * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent standard error of the mean (SEM).

    Techniques Used: In Vitro, Expressing, Cell Culture, Immunocytochemistry, High Content Screening, Concentration Assay

    Summary of in vitro and ex vivo systems to mimic alveolar epithelial and fibroblast damage and dysfunction in the IPF lung. While IPF fibroblasts and bleomycin treated NHLFs exhibit some hallmarks of senescence, these cells do not appear to demonstrate a phenotype of Cdkn2a /p16 ink4a expression, ECM deposition, or secretion of fibrotic mediators such as TIMP1. Rather, the SASP of senescent, aberrant epithelial cells drives a fibrotic phenotype in NHLFs that is consistent with progressive fibrosis. Development of senolytic agents presents an opportunity for therapeutic impact early in disease pathogenesis and with an orthogonal mechanism than the fibroblast targeting standard of care, Nintedanib. Image created with https://www.biorender.com/ .
    Figure Legend Snippet: Summary of in vitro and ex vivo systems to mimic alveolar epithelial and fibroblast damage and dysfunction in the IPF lung. While IPF fibroblasts and bleomycin treated NHLFs exhibit some hallmarks of senescence, these cells do not appear to demonstrate a phenotype of Cdkn2a /p16 ink4a expression, ECM deposition, or secretion of fibrotic mediators such as TIMP1. Rather, the SASP of senescent, aberrant epithelial cells drives a fibrotic phenotype in NHLFs that is consistent with progressive fibrosis. Development of senolytic agents presents an opportunity for therapeutic impact early in disease pathogenesis and with an orthogonal mechanism than the fibroblast targeting standard of care, Nintedanib. Image created with https://www.biorender.com/ .

    Techniques Used: In Vitro, Ex Vivo, Expressing

    Related Articles

    In Vitro:

    Article Title: Modulating in vitro lung fibroblast activation via senolysis of senescent human alveolar epithelial cells.
    Article Snippet: .. Cells, and reagents, and in vitro procedures Primary normal human lung fibroblasts (NHLF) were purchased from Lonza and ATCC, and diseased IPF lung fibroblasts were purchased from BioIVT, Lonza, or ATCC. .. These cells were cultured in fibroblast basal medium supplemented with a low-serum (2%) growth kit (ATCC, Manassas, VA, USA) and 1% penicillinstreptomycin (Thermo Fisher Scientific, Waltham, MA, USA) and expanded to no greater than passage 4.



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    Relative mRNA expression levels of Axl, Tyro3, and Gas6 genes in IPF FBs and HPFs quantified by RT-qPCR. No Mer expression was detected in both fibroblast types. TBP gene was used as housekeeping gene. N = 4.

    Journal: Medicina

    Article Title: Evaluation of TAM Receptor Targeting in Pathophysiology of Idiopathic Pulmonary Fibrosis

    doi: 10.3390/medicina61101837

    Figure Lengend Snippet: Relative mRNA expression levels of Axl, Tyro3, and Gas6 genes in IPF FBs and HPFs quantified by RT-qPCR. No Mer expression was detected in both fibroblast types. TBP gene was used as housekeeping gene. N = 4.

    Article Snippet: IPF fibroblasts (IPF FBs CCL-134, ATCC) were cultured in Ham’s F12K medium supplemented with 10% FBS and 1% penicillin/streptomycin solution, and cells were used between passage 11 and 24.

    Techniques: Expressing, Quantitative RT-PCR