epithelial cell complete medium  (Procell Inc)

Journal: MedComm

Article Title: Nitrate Enhances Gastric Mucosa Defense and Repair Process in Ethanol‐Induced Gastric Ulcer Rats via the Notch–Tff2 Pathway

doi: 10.1002/mco2.70628

Figure Lengend Snippet: Dietary nitrate attenuates ethanol‐induced gastric mucosal hemorrhage and oedema in vivo. (A) Establishment of ethanol‐induced gastric ulcer in rats by intragastric administration of anhydrous ethanol. (B) Flowchart of the animal experimental procedures. (C) Macroscopic appearance of the gastric mucosa in four groups. (D) Quantification of gastric damage expressed as the ulcer index (UI). The UI was calculated as follows: UI = 10 × (ulcerated area/total mucosal area). (E) Representative HE‐stained images of stomach tissues. Scale bar = 50 µm. (F) Histological evaluation of gastric lesions using a microscopic HE score (0–14). The score sums the severity of four features: inflammatory cells (0–3), mucosal edema (0–4), hemorrhage (0–4), and epithelial loss (0–3). (G) Representative CD31 IF staining images in gastric mucosa. Scale bar = 20 µm. (H and I) RT‐qPCR analysis of Ang1 and Et1 mRNA expression in gastric mucosa. Target gene expression was normalized to Gapdh mRNA and expressed as fold change relative to the control group. (J and K) The nitrate levels of the serum and gastric mucosa in the four groups. Quantitative data are expressed as the mean ± SD. * p < 0.05, *** p < 0.001, and ns denotes no significance. HE, hematoxylin–eosin; Nit, nitrate; IF, immunofluorescence; CD31, platelet endothelial cell adhesion molecule‐1; Ang1, angiotensin‐1; Et1, endothelin‐1; RT‐qPCR, real‐time quantitative polymerase chain reaction; DAPI, 2‐(4‐amidinophenyl)‐6‐indolecarbamidine dihydrochloride; SD, standard deviation.

Article Snippet: Primary human gastric mucosal epithelial (CP‐H048) cells and primary human gastric mucosal epithelial cell complete medium (CM‐H048) were purchased from Procell (Wuhan, Hubei, China).

Techniques: In Vivo, Staining, Quantitative RT-PCR, Expressing, Targeted Gene Expression, Control, Immunofluorescence, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: MedComm

Article Title: Nitrate Enhances Gastric Mucosa Defense and Repair Process in Ethanol‐Induced Gastric Ulcer Rats via the Notch–Tff2 Pathway

doi: 10.1002/mco2.70628

Figure Lengend Snippet: Tff2 knockdown abolishes nitrate protection against ethanol‐induced gastric ulcers in vivo. (A) Establishment of Tff2‐KD rat by tail vein injection of recombinant AAV. (B) The timeline of dietary nitrate administration (3 weeks after injection and 7 days before ethanol gavage). (C and D) The macroscopic appearance and ulcer index of the gastric mucosa in Tff2‐KD and scramble groups with ethanol gavage. The UI was calculated as follows: UI = 10 × (ulcerated area/total mucosal area). (E and F) Representative histology images of stomach tissue and a histopathologic score of HE staining in Tff2‐KD and scramble groups. The HE score sums the severity of four features: inflammatory cells (0–3), mucosal edema (0–4), hemorrhage (0–4), and epithelial loss (0–3). Scale bar = 50 µm. (G and H) Representative gastric tissue images of AB–PAS staining and mucin histochemical analysis of Tff2‐KD and scramble groups. The mucin area was expressed as fold change relative to the scramble + EtOH group. Scale bar = 50 µm. Quantitative data are expressed as the mean ± SD. *** p < 0.001, and ns denotes no significance. AAV, adeno‐associated virus; HE, hematoxylin–eosin; AB–PAS, Alcian blue and periodic acid‐Schiff; EtOH, ethanol; KD, knockdown; Nit, nitrate; Tff2; trefoil factor 2.

Article Snippet: Primary human gastric mucosal epithelial (CP‐H048) cells and primary human gastric mucosal epithelial cell complete medium (CM‐H048) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Knockdown, In Vivo, Injection, Recombinant, Staining, Virus

Journal: MedComm

Article Title: Nitrate Enhances Gastric Mucosa Defense and Repair Process in Ethanol‐Induced Gastric Ulcer Rats via the Notch–Tff2 Pathway

doi: 10.1002/mco2.70628

Figure Lengend Snippet: Nitrate promotes migration by TFF2 upregulation in vitro. (A) Images of the scratch healing process of GES‐1 cells in Ibidi culture inserts. Scale bar = 500 µm. (B and C) Quantitative analysis of the migration rate at 24 h and 48 h in (A). (D and E) Representative immunoblotting band of the TFF2 protein and analysis of band gray values. (F) IF staining of pMLC (pink) and DAPI (blue). Scale bar = 40 µm. (G) IF analysis of pMLC with MFI. (H) Images of the scratch healing process of GES‐1 cells transfected with si‐ TFF2 /si‐negative control in Ibidi culture‐inserts. Scale bar = 500 µm. (I and J) Quantitative analysis of the migration rate at 24 h and 48 h. (K) Images of the scratch healing process of GES‐1 cells transfected with si‐ TFF2 /si‐negative control in Ibidi culture‐inserts containing ethanol. Scale bar = 500 µm. (L and M) Quantitative analysis of the migration rate at 24 h and 48 h. Migration rate is quantified by the percentage of closed area to the initial scratch area. Quantitative data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, and ns denotes no significance. TFF2, trefoil factor 2; GES‐1, human gastric epithelial; EtOH, ethanol; Nit, nitrate; Ctrl, control; MLC, myosin light chain 2; pMLC, phosphorylated myosin light chain 2; DAPI, 2‐(4‐amidinophenyl)‐6‐indolecarbamidine dihydrochloride; NC, negative control; SD, standard deviation.

Article Snippet: Primary human gastric mucosal epithelial (CP‐H048) cells and primary human gastric mucosal epithelial cell complete medium (CM‐H048) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Migration, In Vitro, Western Blot, Staining, Transfection, Negative Control, Control, Standard Deviation

Journal: MedComm

Article Title: Nitrate Enhances Gastric Mucosa Defense and Repair Process in Ethanol‐Induced Gastric Ulcer Rats via the Notch–Tff2 Pathway

doi: 10.1002/mco2.70628

Figure Lengend Snippet: Nitrate functions by Notch pathway inhibition positively transcripting TFF2 in vitro. (A) Diagram of the wild‐type and mutant sequences for the three predicted RBPJ binding sites within the 251 bp TFF2 promoter probes. (B) Representative electrophoretic mobility shift assay (EMSA) autoradiograph. Hot probe is the biotin‐labeled wild‐type oligonucleotides of the truncated TFF2 promoter containing the binding motif; Mutant probe is the labeled oligonucleotides sequence with nucleotides mutated. The cold probe is nonlabeled competitive wild‐type probes (100 and 50 that of the concentrations). The shifted bands are indicated by arrows, which suggested the formation of DNA–protein complexes (lane 2, 3, 7). The super shifted bands indicated the formation of DNA–protein–antibody complexes (lane 3, 7). “+” and “−” represent presence and absence, respectively. (C) Relative TFF2 promoter (Full, Mut1, Mut2, and Mut3) luciferase activity was detected by DLR assays in RBPJ overexpressed and normal‐expressed GES‐1 cells. (D) A schematic diagram showing the location of RBPJ putative binding regions on the TFF2 promoter. (E) RT‐qPCR analysis of TFF2 binding site expression of GES‐1 cells. Target site expression in RBPJ‐treated groups was normalized to IgG negative control groups and expressed as fold change relative to the IgG groups. (F) IF staining of NICD (pink) and DAPI (blue). Scale bar = 50 µm. (G) IF analysis of MFI of nuclear NICD in positive cells. (H) Representative immunoblotting band of Notch signaling pathway in GES‐1 cells. (I–K) Analyses of immunoblotting band gray value of (H). (L) RT‐qPCR analysis of TFF2 mRNA expression of DMSO/DAPT treated GES‐1 cells. Target gene expression was normalized to GAPDH mRNA and expressed as fold change relative to the DMSO vehicle group. (M) RT‐qPCR analysis of TFF2 mRNA expression in RBPJ overexpressed and NICD deprived GES‐1 cells. The target gene expression was normalized to GAPDH mRNA and expressed as fold change relative to the vector + DMSO group. (N) RT‐qPCR analysis of TFF2 mRNA expression in RBPJ overexpressed and NICD‐RBPJ overexpressed GES‐1 cells. The target gene expression was normalized to GAPDH mRNA and expressed as fold change relative to the vector1 + vector2 group. (O) Representative image of PLA of NICD–RBPJ proximity. Each red dot represents a positive signal of NICD–RBPJ interaction and nuclei were counterstained with DAPI (blue). Scale bar = 20 µm. Quantitative data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns denotes no significance. Nit, nitrate; EtOH, ethanol; GES‐1, human gastric epithelial; NICD, intracellular structural domain; RBPJ, recombination signal binding protein for immunoglobulin kappa J region; EMSA, electrophoretic mobility shift assay; TFF2, trefoil factor 2; DAPI, 2‐(4‐amidinophenyl)‐6‐indolecarbamidine dihydrochloride; Yhhu‐3792, N2‐(4‐isopropylphenyl)‐5‐(3‐methoxyphenoxy) quinazoline‐2,4‐diamine; Luc, luciferase; SD, standard deviation; DLR, dual‐luciferase report; PLA, proximity ligation assay.

Article Snippet: Primary human gastric mucosal epithelial (CP‐H048) cells and primary human gastric mucosal epithelial cell complete medium (CM‐H048) were purchased from Procell (Wuhan, Hubei, China).

Techniques: Inhibition, In Vitro, Mutagenesis, Binding Assay, Electrophoretic Mobility Shift Assay, Autoradiography, Labeling, Sequencing, Luciferase, Activity Assay, Quantitative RT-PCR, Expressing, Negative Control, Staining, Western Blot, Targeted Gene Expression, Plasmid Preparation, Standard Deviation, Proximity Ligation Assay

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: DOG screening overview. a , Screening overview showing the heatmaps (average of n = 3) of the cell proliferation at 24 h and 72 h for both the MCF10a and MCF7 cells quantified as Ki-67 positive nuclei. The heatmaps corresponding to the cell density and cell area can be found in the supplementary figures. b , Scatter plots showing all data points from one screening (4 DOGs) ranked from low to high. Tissue culture polystyrene (TCP) was used as a control. The 10 % lowest and 10 % highest scoring datapoints are shown on the side and are shown to be significantly different (p < 0.0001). Annotation of black dot with “T” or “L” corresponds to the representative fluorescence images of “Top” and “Low” hits as shown in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Control, Fluorescence

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Top- and low-hit Ki-67-expressing proliferative cells after 72 h of cell culture . Cells were stained for Ki-67 (red), phalloidin (green), and nuclei (blue). Scale bar is 100 μm. White arrows show examples of Ki-67 positive nuclei. The underlying material properties of each image are: MCF10a top hit: S-W 51° 251 MPa, MCF10a low hit: T-S|W 3.8 μm 82° 47 MPa, MCF7 top hit: S-W 63° 42 MPa, MCF7 low hit: T-SW 6 μm 68° 63 MPa.

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Expressing, Cell Culture, Staining

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Scatter plots comparing the effect of each individual physicochemical parameter of the DOG between MCF10a and MCF7 cells. The material parameters (topography, wettability, and stiffness on the same data points, shown with the corresponding color scale) were shown on the biological output (cell proliferation, % Ki-67 positive cells) for the MCF10a vs. the MCF7 cells. Similar plots showing the scatter plots for cell density and cell area can be found in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques:

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: ROIs were chosen that lead to a higher proliferation after 72 h for MCF10a than MCF7 cells. a , Regions of interest (ROIs) are chosen as points furthest away from the dashed line. In orange and blue, points are indicated that show an individual statistical difference (P < 0.05) when comparing the proliferation percentage of MCF7 vs. MCF10a. Orange or blue dots indicate an increase for either MCF7 or MCF10a proliferation, respectively. b , Points from a were grouped and show a significant different in both MCF10a and MCF7 Ki-67 positive cells percentage (p < 0.0001). c , Individual DOG values for each ROI chosen in a are shown. Corresponding fluorescence images for each ROI can be found in .

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Fluorescence

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: ROIs were chosen from the screening as “positive” when MCF10a Ki-67 % > MCF7 Ki-67 % (labelled as Pos1, Pos2, Pos3, shown in blue) and “negative” when MCF7 Ki-67 % > MCF10a Ki-67 % (labelled as Neg1, Neg2, Neg3, shown in orange). a , Cell experiments were repeated on translational (labelled as Trans) substrates with those specific material properties and compared to the proliferation (% Ki-67 positive cells after 72 h of cell culture) rates from the screening (Screen). b , Cell density after 72 h was compared between the screening and translation findings. c , Fluorescence images of two example ROIs, Pos1 and Neg1. Cells were stained for Ki-67 (red), phalloidin (green), and DAPI (blue). Corresponding fluorescence images for the other ROIs can be found in . The scale bar is 100 μm.

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques: Cell Culture, Fluorescence, Staining

Journal: Bioactive Materials

Article Title: Harnessing the power of physicochemical material property screening to direct breast epithelial and breast cancer cells

doi: 10.1016/j.bioactmat.2025.04.003

Figure Lengend Snippet: Proliferation quantified as % Ki-67 positive cells after 72 h for both MCF7 and MCF10a cells compared between the translational cell experiments on translated ROIs for both low (shown in blue, 1000 cells cm −2 ) and high (shown in orange, 5000 cells cm −2 ) seeding density. a , Individual values for each ROI compared between low and high seeding density. b , All low and high seeding density datapoints from a were grouped and compared for statistical difference (n.s.: p > 0.05, ∗: p < 0.05).

Article Snippet: Normal breast epithelial cells (MCF10a, Aldrich) were maintained in MCF10a Complete Medium (EP-ML-0525, Elabscience) containing 5 % horse serum, 20 ng mL −1 epidermal growth factor (EGF), 0.5 μg/mL hydrocortisone, 10 μg/mL insulin, 1 % non-essential amino acid solution (NEAA), and 1 % penicillin/streptomycin.

Techniques:

Journal: Frontiers in Immunology

Article Title: CYP1B1 knockout enhanced IFN-γ production is required but not sufficient for protection of cigarette smoke-exposed mice against lethal influenza virus infection

doi: 10.3389/fimmu.2025.1600025

Figure Lengend Snippet: CYP1B1 is induced by IAV and CSE in BciNS1 cells, and TMS protects airway epithelial cells from cell death induced by IAV infection. BciNS1 cells were incubated with 0% (Mock) and 5% CSE for 24h. For IAV infection, the cells were exposed to IAV PR8 at an MOI of 0.1 for 24h. (A) CYP1B1 mRNA levels were assessed by qRT-PCR and normalized to β-actin. (B) CYP1B1 protein expression in BciNS1 cells was determined by western blot. Data are expressed as means ± SD. * denotes a significant difference compared to the mock group ( P < 0.05 by one-way ANOVA with Tukey test for multiple comparisons, n = 3.). (C) LDH activity in the supernatant and cell extract was measured using an LDH Cytotoxicity Assay Kit (BioVision Research Products). Cytotoxicity is expressed as the percentage of supernatants released LDH to total (supernatant + cell) LDH. * denotes a significant difference between the two groups ( P < 0.05 by one-way ANOVA with Tukey test for multiple comparisons, n = 3).

Article Snippet: Cells were resuspended to 5 × 10 5 cells/ml in complete Bronchial Epithelial Cell Growth Medium (BEGM; Lonza Group Ltd.), were seeded into collagen-coated tissue culture plates (Bio-Coat, BD Biosciences) at a density of 1 × 10 5 cells/cm 2 , and were propagated in an incubator at 37°C in 5% CO 2 .

Techniques: Infection, Incubation, Quantitative RT-PCR, Expressing, Western Blot, Activity Assay, LDH Cytotoxicity Assay