Dataset Supporting the Protocol for Mucus Producing Gut on a Chip Construction

Overview: This dataset contains 189 image files, summarizing data from gut on chip (GoC) development and is comprised of brightfield microscopy, confocal microscopy and brightfield microscopy with mucin-specific staining. Additionally, primary data on cell viability using CCK-8 assay are summarized and included in this dataset. Both brightfield and confocal microscopy images confirms successful model development. 

Structure of the dataset: The dataset is split into three main folders that correspond to:

Brightfield microscopy images for both 7:3 and 9:1 seeding ratios of Caco-2 and HT29-MTX-E12 co-culture. Microscopy was performed after 1, 2, 4, 6 and 8 days of cells growth. All image files are renamed according to the following template: GOC_[ratio]_[day]_[numerator], simultaneously subfolder structure follows the same pattern.

Confocal microscopy images were also performed for both 7:3 and 9:1 seeding ratios of Caco-2 and HT29-MTX-E12 co-culture. At the end of the experiment (day 8) cells in the chip were fixed with formaldehyde and staining was performed using DAPI, F-actin, Muc5AC, VIL1 and ZO1 as fluorescent markers. Similarly, all confocal image files are renamed according to the following template: GOC_[ratio]_[marker]_[numerator], simultaneously subfolder structure follows the same pattern.

Brightfield microscopy images for cells fixed (with Carnoy’s fixative) and stained with Alcian Blue / PAS (Periodic-Schiff acid) dye were performed also for both (i.e., 7:3 and 9:1) co-culture variants at the end of the experiment. Hence, image files are renamed according to the following template: GOC_[ratio]_mucin_[numerator] and separated in two subfolders as per cell seeding ratio.

Additionally, text document summarizing data on cell viability (using CCK8 viability assay) and membrane integrity (using Lucifer Yellow for permeability testing) is included in the dataset.

Acknowledgement: The work presented in this paper was supported by the Latvian Council of Science Fundamental and Applied Research Project 'Research of microbiota-derived extracellular vesicle role on breast cancer by using gut-breast cancer axis on a chip', project No: LZP-2021/1-0041.