caco 2 cells  (ATCC)

Journal: Poultry Science

Article Title: Houttuynia cordata extract protects against Salmonella infection by targeting type III secretion system 1

doi: 10.1016/j.psj.2026.106937

Figure Lengend Snippet: HCE suppresses the adhesion and invasion of Salmonella in Caco-2 cells. A CCK-8 assay (A) was conducted to evaluate the cytotoxicity of HCE in Caco-2 cells. The effects of HCE on the adhesion (B, D) and invasion (C, E) of ST (B-C) and SP (D-E) to Caco-2 cells were examined at an MOI of 100 (n = 5). Immunofluorescence microscopy was performed to visualize the inhibitory effect of HCE (150 µg/mL) on the invasion of ST (F) and SP (G) in Caco-2 cells. Caco-2 cells infected with the ST Δ invA strain, which was used as a negative control because of its T3SS-1 deficiency showed significantly reduced invasion. ST, Salmonella Typhimurium. SP, Salmonella Pullorum . HCE, Houttuynia cordata extract.

Article Snippet: Caco-2 cells (American Type Culture Collection, ATCC, USA) were cultured in DMEM and seeded into culture plates.

Techniques: CCK-8 Assay, Immunofluorescence, Microscopy, Infection, Negative Control

Journal: Materials Today Bio

Article Title: Biomimetic villi-crypt scaffold-on-chip with tunable mechanical properties for intestinal epithelium modeling

doi: 10.1016/j.mtbio.2026.103134

Figure Lengend Snippet: Villi-crypt in vitro model. Resazurin reduction assay performed (a) over the Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds in the presence and absence of gelatin to aid cell attachment ( n = 3 scaffolds for each condition). (b) Resazurin reduction assay performed for long-term cultures in the absence of gelatin (two independent experiments, n = 5 scaffolds for each condition of each experiment). Data are expressed as mean ± standard deviation. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. (c) Phalloidin (green) and DAPI (blue) fluorescence microscopy of Caco-2 cells grown for 21 days on the two compositions tested at two different magnifications. (c i ) Top and (c ii ) section view of the scaffolds. Scale bars represent 500 μm and 100 μm. Objectives are 4 × and 10 × , respectively. (d) Confocal images of Villi-(R)-Crypt scaffold cultured with the Caco-2 cells. (d i ) Schematic of the different confocal planes acquired. (d ii ) and (d iii ) details at two different magnifications of villi sections, scaffold baselines and crypt base; displaying the homogeneous covering of the scaffold and junctions formed between cells ( V = villi, C = crypt). Scale bars represent 200 μm (d ii , 10 × objective) and 50 μm (d iii , 20 × objective). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Cell cultures over scaffolds: Human epithelial cell line Caco-2 (HTB-37, ATCC, USA) was cultured in 75 cm 2 tissue culture polystyrene flasks in complete DMEM high glucose (Sigma Aldrich, pc D6546) supplemented with 1% penicillin-streptomycin (Sigma Aldrich, pc P0781), 1% L-glutamine (Sigma Aldrich, pc G7513) and 10% FBS (Invitrogen, USA) in a humidified incubator set at 37 °C with 95% air and 5% CO 2 .

Techniques: In Vitro, Cell Attachment Assay, Standard Deviation, Fluorescence, Microscopy, Cell Culture

Journal: Materials Today Bio

Article Title: Biomimetic villi-crypt scaffold-on-chip with tunable mechanical properties for intestinal epithelium modeling

doi: 10.1016/j.mtbio.2026.103134

Figure Lengend Snippet: (a) Fold change in gene expression with respect to undifferentiated cells and (b) quantification of produced proteins. Caco-2 cells were cultured for 14 and 21 days on the two compositions tested ( n = 3 scaffolds for each condition). Data are expressed as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Cell cultures over scaffolds: Human epithelial cell line Caco-2 (HTB-37, ATCC, USA) was cultured in 75 cm 2 tissue culture polystyrene flasks in complete DMEM high glucose (Sigma Aldrich, pc D6546) supplemented with 1% penicillin-streptomycin (Sigma Aldrich, pc P0781), 1% L-glutamine (Sigma Aldrich, pc G7513) and 10% FBS (Invitrogen, USA) in a humidified incubator set at 37 °C with 95% air and 5% CO 2 .

Techniques: Gene Expression, Produced, Cell Culture, Standard Deviation

Journal: Materials Today Bio

Article Title: Biomimetic villi-crypt scaffold-on-chip with tunable mechanical properties for intestinal epithelium modeling

doi: 10.1016/j.mtbio.2026.103134

Figure Lengend Snippet: Venn diagram showing the distribution of proteins identified in Villi-(F)-Crypt and Villi-(R)-Crypt scaffolds cultured with Caco-2 cells for the period of 14 and 21 days. The core proteins are shared across all conditions, while the exclusive proteins represent those uniquely identified in specific conditions. The 10 most abundant proteins are highlighted.

Article Snippet: Cell cultures over scaffolds: Human epithelial cell line Caco-2 (HTB-37, ATCC, USA) was cultured in 75 cm 2 tissue culture polystyrene flasks in complete DMEM high glucose (Sigma Aldrich, pc D6546) supplemented with 1% penicillin-streptomycin (Sigma Aldrich, pc P0781), 1% L-glutamine (Sigma Aldrich, pc G7513) and 10% FBS (Invitrogen, USA) in a humidified incubator set at 37 °C with 95% air and 5% CO 2 .

Techniques: Cell Culture

Journal: Clinical & Translational Immunology

Article Title: Aberrant intra‐epithelial lymphocytes cause enterocyte cell death in refractory celiac disease by CD103 ‐β7‐receptor‐mediated granzyme‐B degranulation which can be restored by etrolizumab

doi: 10.1002/cti2.70099

Figure Lengend Snippet: Granzyme‐B is the key cell death mediator in enterocyte‐induced cell death and is only secreted by aberrant intra‐epithelial T‐lymphocyte (IEL) in the presence of enterocytes. (a) Cytoplasmic localisation of granzyme‐B granules in refractory celiac disease type II (RCDII) cells P2 using immunofluorescence (red, granzyme‐B; blue, DAPI nucleus staining, 100× magnification). (b) Expression of degranulation marker CD107a on RCDII cell lines P1 and P2 in the absence and presence of epithelial Caco2 cells after 4 h of incubation. (c) CD107a expression on aberrant IEL of a duodenal biopsy from a representative RCDII patient with villous atrophy after 4 h of incubation; left panel, isotype‐matched control. (d) Granzyme‐B secretion by RCDII cell lines in the absence and presence of enterocyte cell line Caco2 after 6 h of incubation. (e) RCDII cell lines P1 and P2 induce killing of epithelial Caco2 cells. The control cell line SUDHL4 showed no cytotoxicity against the Caco2 cells. (f) Enterocyte cell death by RCDII cells incubated with increasing concentrations of degranulation blocker hydroxychloroquine sulphate (HCQ). (g) Killing of intestinal Caco2 cells by RCDII cells in the presence of increasing concentrations of the granzyme‐B inhibitor Z‐AAD‐CH2Cl. For cell death assays, cytotoxicity was measured after 16 h of co‐incubation at an effector:target ratio 2:1. Cell experiments were done in triplicate. *** P ≤ 0.001, unpaired t ‐test, results are shown as mean + sem.

Article Snippet: The intestinal epithelial cell line Caco2 was obtained from the American Type Culture Collection (ATCC) and cultured with DMEM medium (BioWhittaker) containing 10% FBS (GE Healthcare Life Sciences) and 100 IU penicillin/100 μg/mL streptomycin (1% P/S) at 37°C.

Techniques: Immunofluorescence, Staining, Expressing, Marker, Incubation, Control

Journal: Clinical & Translational Immunology

Article Title: Aberrant intra‐epithelial lymphocytes cause enterocyte cell death in refractory celiac disease by CD103 ‐β7‐receptor‐mediated granzyme‐B degranulation which can be restored by etrolizumab

doi: 10.1002/cti2.70099

Figure Lengend Snippet: Aberrant intra‐epithelial T‐lymphocyte (IEL) demonstrate upregulated expression of CD103 and require cell–cell binding to induce enterocyte killing. (a) Refractory celiac disease type II (RCDII) cell‐induced cytotoxicity of Caco2 cells in the presence of a transwell system. Cell death was measured after 16 h of co‐incubation at an effector:target ratio 2:1. (b) Degranulation by RCDII cells in the presence of a transwell system. CD107a expression was measured after 4 h of co‐incubation with Caco2 cells at an effector:target ratio 2:1. (c) NKG2D expression on aberrant IEL of RCDII patients and patients with celiac disease (CD) on gluten‐free diet (GFD), using flow cytometry analysis. Activated CD8+ T cells served as positive control. (d) Representative histograms of NKG2D expression on RCDII cell lines P1 and P2 using flow cytometry; grey shaded peak, isotype‐matched control. (e) Killing of epithelial cells Caco2 by RCDII cell line P1 in the presence of 20 μg/mL NKG2D‐blocking mAb or isotype control mAb. (f) CD103 expression on aberrant IEL of RCDII patients and patients with CD on GFD, using flow cytometry analysis. (g) Follow‐up of CD103 expression on aberrant IEL from a representative RCDII patient, showing persistent villous atrophy after first‐line treatment (cladribine; non‐responding) and complete mucosal recovery after second‐line treatment (autologous stem cell transplantation; responding). t = 0 at diagnosis and start treatment, t = 1 is 3 months after first‐line treatment, t = 2 and t = 3 is 3, respectively, 6 months after stem cell transplantation. (h) Histograms of CD103 expression on RCDII cell lines P1 and P2 using flow cytometry analysis; grey shaded peak, isotype‐matched control. Cell experiments were done in triplicate. * P ≤ 0.05, *** P ≤ 0.001, unpaired t ‐test, results shown as mean + sem.

Article Snippet: The intestinal epithelial cell line Caco2 was obtained from the American Type Culture Collection (ATCC) and cultured with DMEM medium (BioWhittaker) containing 10% FBS (GE Healthcare Life Sciences) and 100 IU penicillin/100 μg/mL streptomycin (1% P/S) at 37°C.

Techniques: Expressing, Binding Assay, Incubation, Flow Cytometry, Positive Control, Control, Blocking Assay, Transplantation Assay, Biomarker Discovery

Journal: Clinical & Translational Immunology

Article Title: Aberrant intra‐epithelial lymphocytes cause enterocyte cell death in refractory celiac disease by CD103 ‐β7‐receptor‐mediated granzyme‐B degranulation which can be restored by etrolizumab

doi: 10.1002/cti2.70099

Figure Lengend Snippet: Aberrant intra‐epithelial T‐lymphocyte (IEL)‐enterocyte binding via CD103 induces granzyme‐B‐mediated enterocyte cell death. (a) Killing of Caco2 epithelial cells by refractory celiac disease type II (RCDII) cell lines in the presence of 10 μg/mL CD103‐blocking mAb or isotype control. Cell death was measured after 16 h of co‐incubation at an effector:target ratio 2:1. (b) Degranulation by RCDII cell lines, measured by CD107a expression, in the presence of 10 μg/mL CD103‐blocking mAb or the matching isotype control. Degranulation was measured after 4 h of co‐incubation with Caco2 cells at an effector:target ratio 2:1. (c) Secretion of granzyme‐B by RCDII cell lines co‐incubated with Caco2 cells in the presence of 10 μg/mL CD103‐blocking mAb compared to the isotype control. Secretion was measured after 6 h of co‐incubation at an effector:target ratio 2:1. (d) Upper left picture: small intestinal organoids; upper right picture: attachment of RCDII cells to an organoid (blue arrow); lower left picture: RCDII cells induce killing of organoids (red arrows); lower right picture: in the presence of 10 μg/mL CD103‐blocking mAb RCDII–induced organoid cell death is evidently reduced, illustrated by the presence of viable organoids (green arrows). (e) Induction of organoid cell death by RCDII cells P2 in the presence of 10 μg/mL CD103‐blocking antibody or an isotype control, measured by cell count via microscopy. Killing was measured after 24 h of co‐incubation at an effector:target ratio 50:1. Figure , upper pictures 25× magnification, lower pictures 10× magnification, using an Olympus microscope. Cell experiments for Figure performed in duplicate, other cell experiments performed in triplicate. *** P ≤ 0.001, unpaired t ‐test, results shown as mean + sem.

Article Snippet: The intestinal epithelial cell line Caco2 was obtained from the American Type Culture Collection (ATCC) and cultured with DMEM medium (BioWhittaker) containing 10% FBS (GE Healthcare Life Sciences) and 100 IU penicillin/100 μg/mL streptomycin (1% P/S) at 37°C.

Techniques: Binding Assay, Blocking Assay, Control, Incubation, Expressing, Cell Characterization, Microscopy

Journal: Clinical & Translational Immunology

Article Title: Aberrant intra‐epithelial lymphocytes cause enterocyte cell death in refractory celiac disease by CD103 ‐β7‐receptor‐mediated granzyme‐B degranulation which can be restored by etrolizumab

doi: 10.1002/cti2.70099

Figure Lengend Snippet: Etrolizumab inhibits granzyme‐B secretion and restores enterocyte viability. (a) Histograms of β7 expression on refractory celiac disease type II (RCDII) cell lines P1 and P2; grey shaded peak, isotype‐matched control. (b) Caco2 cell death by RCDII cells in the presence of 50 μg/mL etrolizumab or isotype control. Cell death was determined after 16 h of co‐incubation at an effector:target ratio 2:1. (c) Degranulation by RCDII cell lines in the presence of 50 μg/mL etrolizumab or the matching isotype control. Degranulation was measured after 4 h of co‐incubation with Caco2 cells at an effector:target ratio 2:1. (d) Secretion of granzyme‐B by RCDII cell lines co‐incubated with enterocyte Caco2 cells in the presence of 50 μg/mL etrolizumab compared to the isotype control. Secretion was measured after 6 h of co‐incubation at an effector:target ratio 2:1. (e) Upper picture: RCDII cells attach to the organoid surface and cause membrane disruption (red arrow), inducing organoid cell death; lower picture: the attachment of RCDII cells to organoids and subsequent organoid cell death is decreased in the presence of 50 μg/mL etrolizumab antibody. (f) Induction of organoid cell death by RCDII cells P2 in the presence of 50 μg/mL etrolizumab or an isotype control, measured by cell count via microscopy. Killing was measured after 24 h of co‐incubation at an effector:target ratio 20:1. Figure , pictures 10× magnification, using an Olympus microscope. Cell experiments were done in triplicate. *** P ≤ 0.001, unpaired t ‐test, results shown as mean + sem.

Article Snippet: The intestinal epithelial cell line Caco2 was obtained from the American Type Culture Collection (ATCC) and cultured with DMEM medium (BioWhittaker) containing 10% FBS (GE Healthcare Life Sciences) and 100 IU penicillin/100 μg/mL streptomycin (1% P/S) at 37°C.

Techniques: Expressing, Control, Incubation, Membrane, Disruption, Cell Characterization, Microscopy

Journal: Aging Cell

Article Title: Age‐Associated Impairment of Paneth Cells Driven by microRNA ‐152 Promotes Intestinal Epithelial Vulnerability to Pathological Stress

doi: 10.1111/acel.70542

Figure Lengend Snippet: MiR‐152 disrupts mitochondrial function by inhibiting PHB1. (A) MiR‐152 overexpression in IECs. MiR‐152 mimic or scramble control was transfected in Caco‐2 cells for 48 h before miR assay to examine the levels of miR‐152. Values are means ± SEM ( n = 5). * p < 0.05 compared with Scramble. (B) Mitochondrial respiration in cells described in (A) examined by Seahorse assay. Values are means ± SEM ( n = 3). * p < 0.05 compared with Scramble. (C) Calculated spare respiration capacity. (D) Immunoblots of mitochondrial proteins in cells described in (A). (E) Biotin labeled miR‐152 or scrambled RNA transfection in IECs. Caco‐2 cells were transfected 48 h before qPCR to examine the levels of miR‐152. Values are means ± SEM ( n = 3). * p < 0.05 compared with biotin labeled Scramble. (F) Pulldown assay to determine the miR‐152 bound mRNAs (left) and Phb1 mRNA levels in input RNA samples (right). Values are means ± SEM ( n = 3). * p < 0.05 compared with biotin labeled Scramble. (G) Levels of luciferase activity of Phb1 3′UTR reporters with or without miR‐152 binding site (BS). Top, luciferase vectors; bottom, luciferase activity in IECs with or without miR‐152 overexpression. (H) Mitochondrial function assay in IECs transfected with either control, or miR‐152 mimic, or miR‐152 with PHB1 overexpression. Top, MitoTracker Green intensity; bottom, superoxide production. Values are means ± SEM ( n = 3). * p < 0.05 compared with Scramble control. (I) Imageflow cytometric analysis of Lysozyme positive cell population in aging human intestinal organoids with transfection of control oligonucleotides, miR‐152 inhibitor, or miR‐152 and PHB1 silencer. n > 5000 cells/group. Experiments were repeated 3 times and showed similar results.

Article Snippet: Human colorectal carcinoma Caco‐2 cells were purchased from American Type Culture Collection and maintained under standard culture conditions (Yu et al. ).

Techniques: Over Expression, Control, Transfection, Western Blot, Labeling, Luciferase, Activity Assay, Binding Assay, Functional Assay