Journal: Cells

Article Title: Hydrogel-Integrated Millifluidic Systems: Advancing the Fabrication of Mucus-Producing Human Intestinal Models

doi: 10.3390/cells13131080

Figure Lengend Snippet: Transfer of the experimental approach to undifferentiated organoid-derived ISCs from the human terminal ileum cultured in expansion media. ( A ) Representative confocal fluorescent images of ISC monolayers cultured in static conditions on Matrigel TM -coated insert membranes and hydrogels at the onset of confluency (Day 0). IF staining for proliferative marker KI-67 (yellow), stem cell marker LGR5 (green), and progenitor cell marker OLFM4 (red) with nuclei counterstain (blue). For fluorescent staining, samples are collected from three independent runs (n = 3) and presented as xy projections of their z-stacks, along with their xz/yz projections. ( B ) Representative IF images of ISCs on days 5–7 cultured in expansion media under static and dynamic conditions, stained for the same markers used in ( A ). For fluorescent staining, samples are collected from three independent runs (n = 3) and presented as xy projections of their z-stacks, along with their xz/yz projections.

Article Snippet: The substrates are typically coated with a biomimetic thin layer of extracellular matrix (ECM)-derived proteins, such as growth factor-reduced Matrigel TM , collagen, or laminin, to support ISC adhesion and growth.

Techniques: Derivative Assay, Cell Culture, Staining, Marker

Journal: Cells

Article Title: Hydrogel-Integrated Millifluidic Systems: Advancing the Fabrication of Mucus-Producing Human Intestinal Models

doi: 10.3390/cells13131080

Figure Lengend Snippet: Time-dependent quantification of neutral (PAS assay) and acidic (AB assay) mucins produced by ISCs during static and dynamic culture on Matrigel TM -coated insert membranes and dSIS-MA (1.5 wt%) hydrogels for estimation of the amount of secreted mucus via a standard calibration curve using bovine submaxillary gland mucins. ( A ) Amount of mucins shed to the culture media by intestinal monolayers on a substrate area of approximately 3.4 cm 2 . Samples were taken from the apical supernatant in static cultures and the media reservoir in dynamic cultures and normalized accordingly. ( B ) Amount of cell- or membrane-bound mucins normalized to the DNA content of the monolayer. Data are presented as mean ± SD for independent runs n = 3, where “*” indicates statistical significance ( p < 0.05) and “n.s.” indicates non-significance of all samples performed by the Kruskal–Wallis test, followed by Dunn’s test.

Article Snippet: The substrates are typically coated with a biomimetic thin layer of extracellular matrix (ECM)-derived proteins, such as growth factor-reduced Matrigel TM , collagen, or laminin, to support ISC adhesion and growth.

Techniques: PAS Assay, Produced, Membrane