pore transparent transwell inserts  (Greiner Bio)

Journal: Journal of Functional Biomaterials

Article Title: “Attractive” Treatment for Abdominal Aortic Aneurysm Repair: Magnetic Localization of Silk-Iron Packaged Extracellular Vesicles

doi: 10.3390/jfb16110395

Figure Lengend Snippet: SIPEs released more ASC-EVs in 24 h compared to 6 h and had higher signal in the Cy5 channel compared to SIMPs. After co-culture with transwell inserts, SIPEs had more Cy5 signal (and therefore ASC-EVs) at 24 h ( B ) compared to that at 6 h ( A ). SIMPs had little to no signal in the Cy5 channel at both 6 h ( C ) and 24 h ( D ).

Article Snippet: For treatments, 0.5 mL of experimental media was added into treatment wells before adding 0.4 μm pore transparent transwell inserts (Greiner, Monroe, NC, USA, 662641) with 150 μL of treatment.

Techniques: Co-Culture Assay

Journal: Journal of Virology

Article Title: Abortive PDCoV infection triggers Wnt/β-catenin pathway activation, enhancing intestinal stem cell self-renewal and promoting chicken resistance

doi: 10.1128/jvi.00137-25

Figure Lengend Snippet: Effect of PDCoV infection on the intestinal barrier integrity of chicken and pig intestinal organoid monolayers. (A–D) mRNA levels of goblet cell-secreted mucin ( MUC2 ) and tight junction proteins ( ZO1 , occludin , and claudin- 1) from PDCoV-infected chicken and pig intestinal organoid monolayers. (E) Illustration of the measurement of electrical resistance across the organoid monolayers in Transwell plates. TEER of (F) chicken and (G) porcine organoid monolayers over 48 h. Viral replication in (H) chicken and (I) pig intestinal organoid monolayers was quantified by RT-qPCR at 24 hpi. (MOI = 0.1). TEER values were calculated from PDCoV-infected (J) chicken and (K) pig intestinal organoid monolayers. (L) Schematic of the Transwell co-culture system. (M) Cytopathic effect of PDCoV-infected ST cells (in the lower chamber) as observed by light microscope (white arrows). Scale bar = 50 µm. (N) Viral replication in ST cells from panel M was quantified by RT-qPCR. ** P < 0.01. ns, not significant.

Article Snippet: Well-developed intestinal organoids were dissociated into single cells, and 2.5 × 10 4 cells were then seeded on 24-well transparent Transwell inserts (0.4 μm pore size, 6.5 mm diameter; Corning).

Techniques: Infection, Quantitative RT-PCR, Co-Culture Assay, Light Microscopy

Journal: Frontiers in Immunology

Article Title: Targeting stemness pathways modulates macrophage polarization and reprograms the tumor microenvironment

doi: 10.3389/fimmu.2025.1513404

Figure Lengend Snippet: CRC-induced macrophage polarization and its association with specific cytokine production signatures. (A) Heatmap showing the marker expression on M0 macrophages after 48-hour transwell co-culture with CRC cell lines of different molecular subtype (CMS 1-4). Macrophage polarization was assessed by flow cytometry using specific M1 (CD86, CD80, CD274, MHC II, CD11c) and M2 (CD163, CD206) markers. Data are presented as modified z-scores of MFI, with red indicating high expression and blue indicating low expression. (B) Hierarchical clustering heatmap of cytokine and chemokine production profiles from CRC cell lines, quantified using semi-quantitative membrane-based cytokine array. The relative protein expression in cell lysates is shown as modified z-scores, with red indicating high expression and blue indicating low expression. (C–N) Correlation plots showing cancer cell cytokine production (X axis, SI values) against macrophage marker expression (Y axis, MFI values). Pearson correlation coefficients (r) are displayed in each plot, with shaded areas representing the 95% confidence interval. All shown correlations were statistically significant (two-sided p < 0.05, Pearson correlation test), indicating strong positive or negative associations between specific cytokines and macrophage polarization markers. MFI – median fluorescence intensity, CMS – consensus molecular subtype, SI – signal intensity, a.u. – arbitraty units.

Article Snippet: Twenty-four hours prior to initiating co-culture, 2 × 10^5 colorectal cancer (CRC) cells were seeded into six-well plates containing 0.4 μm pore transparent PET transwell inserts (Sarstedt) in supplemented RPMI medium.

Techniques: Marker, Expressing, Co-Culture Assay, Flow Cytometry, Modification, Membrane, Fluorescence

Journal: Frontiers in Immunology

Article Title: Targeting stemness pathways modulates macrophage polarization and reprograms the tumor microenvironment

doi: 10.3389/fimmu.2025.1513404

Figure Lengend Snippet: Effects of stemness inhibitors on stemness and immunomodulatory surface marker expression in colon cancer cells. Stemness inhibitors affect the phenotype of cancer cells. (A, B) Heatmap illustrating the changes in surface marker expression following monotherapy and combination therapy with stemness inhibitors (salinomycin, SB-431542, JIB-04, and napabucasin) in HCT116 and SW620 cells. (C, D) The effects of co-culture with M0-like macrophages on the expression of surface markers in HCT116 and SW620 cell lines treated with the same inhibitors. Transwell co-cultures were conducted for 48 hours, with medium supplemented with 0.5 µM salinomycin, 10 µM SB-431542, 0.01 µM JIB-04, 0.01 µM napabucasin, and a combination of these compounds. The expression of stemness (CD44, CD133, ESA) and immunomodulation (MHC I, CD274) markers was measured with flow cytometry. Log2 transformation was applied to the data, and color intensity represents changes in surface marker median fluorescence intensity, with red indicating an increase (higher log2 value) and blue indicating a decrease (lower log2 value) relative to control. Data are presented as medians from four independent experiments. Statistical significance compared to control samples is denoted by an asterisk (*) where p < 0.05.

Article Snippet: Twenty-four hours prior to initiating co-culture, 2 × 10^5 colorectal cancer (CRC) cells were seeded into six-well plates containing 0.4 μm pore transparent PET transwell inserts (Sarstedt) in supplemented RPMI medium.

Techniques: Marker, Expressing, Co-Culture Assay, Flow Cytometry, Transformation Assay, Fluorescence, Control

Journal: bioRxiv

Article Title: SSAO-mediated decrease of endothelial BDNF release affects neuronal GluA1 and PSD95 expression: a peripheral mechanism inducing NVU and CNS disturbances

doi: 10.1101/2025.03.04.641410

Figure Lengend Snippet: Neurovascular unit (NVU) in vitro model A, Extended experimental timeline. Upper arrow represents neuron-glia mixed culture timeline, indicated as days in vitro (DIV). Medium changes needed at each time point are indicated. Middle arrow represents hCMEC/D3 endothelial cell line culture timeline. Seeding, confluence, and deprivation steps are indicated and were synchronized to mixed culture development. Day 0 is considered the day when cells achieve confluence and after this, time of this culture is indicated as days after confluence (DAC). Lower arrow represents the timeline once co-culture system is set up, and time is indicated as days in vitro since co-culture (DIV - CC). Treatments are indicated the day they were performed after setting up the co-culture. B, Co-culture experimental scheme with representative phase contrast and immunocytochemistry images from WT endothelial cells (Occ: Occludin, ZO-1: Zonula occludens, H: Hoechst) and neuron-glia mixed culture (GluA1, MAP2) before setting up the co-culture. According to the time line in A , WT endothelial cells seeded in the transwell device (TW) were placed inside the neuron-glia mixed culture well. Both parts can communicate through the semipermeable membrane at the bottom of the transwell.

Article Snippet: Endothelial cells were kept in an incubator at 37°C with a humidified atmosphere with 5% CO 2 . hCMEC/D3 cells were seeded at 2 × 10 5 cells/mL and, when transwell inserts (Tw: Transwell polyethylene terephthalate transparent membrane inserts, pore size 0.4μm; Sarstedt) were used, they were additionally coated with 1% fibronectin (Sigma) in Hanks’ Balanced Salt Solution (HBSS; ThermoFisher Scientific).

Techniques: In Vitro, Co-Culture Assay, Immunocytochemistry, Membrane