Journal: Materials Today Bio
Article Title: Thermoreversible cell-derived extracellular matrix only hydrogel (CEOgel): Development, characterization, and applications
doi: 10.1016/j.mtbio.2026.103040
Figure Lengend Snippet: Biocompatibility and application of CEOgel in vitro . (A) HUVEC, UCMSC, and BJ cells were cultured on CEOgel-coated plates for 24 h and stained with Calcein-AM and EthD-1 to visualize live and dead cells under a fluorescence microscope. (scale bars = 500 μm) (B) Cell viability was quantified by CCK-8 assay after 72 h of indirect co-culture with CEOgel in a transwell system; untreated cells served as controls. (C) The proliferation of HUVEC, UCMSC, and BJ cells was measured by CCK-8 assay after 7 days of incubation with CEOgel. (D) TNF-α secretion from RAW 264.7 macrophages was quantified after 6 h and 24 h of incubation with CEOgel; untreated and LPS-treated cells served as negative and positive controls, respectively. (E) Images show CEOgel before and after 7 days of incubation in cell culture medium. (F) Calcein-AM staining visualized tube formation of HUVECs encapsulated in CEOgel after 1 and 3 days of culture. (scale bars = 200 μm) (G) cENF solutions at 20 and 27 mg/mL were loaded into microfluidic devices, where in situ gelation formed stable CEOgels. (scale bars = 200 μm) (H) Confocal microscopy captured RFP-HUVEC vascular network formation within CEOgel (20 mg/mL) under flow conditions after 24 and 48 h of culture. (scale bars = 200 μm) (I) Quantitative analysis evaluated vascular network parameters, including the number of branch points, meshes, nodes, and total length. (J) Live/dead assays assessed cell viability of CRC organoids in CEOgel at day 2 and 9. (scale bars = 100 μm) (K) Bright field images show colorectal cancer (CRC) organoids grown in CEOgel at day 5 and 10. (scale bars = 100 μm) (L) WST-8 assays quantified CRC-organoid proliferation in CEOgel. Data are presented as mean ± SD; ∗∗ p < 0.01, ∗∗∗ p < 0.001, n.s. = not significant ( p > 0.05); n = 3. (M) Immunofluorescence staining revealed expression of the proliferation marker, Ki-67 and the epithelial adhesion marker, E-cadherin in CRC organoids cultured in 33 mg/mL CEOgel. (scale bars = 50 μm).
Article Snippet: The effect of 3D culture in CEOgel on cellular proliferation was assessed using the water-soluble tetrazolium salt WST-8 (Quanti-MaxTM WST-8 Cell Viability Assay Kit, Biomax, Gyeonggi-do, Korea).
Techniques: In Vitro, Cell Culture, Staining, Fluorescence, Microscopy, CCK-8 Assay, Co-Culture Assay, Incubation, In Situ, Confocal Microscopy, Immunofluorescence, Expressing, Marker
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