Detailed survey of an in vitro intestinal epithelium model by single-cell transcriptomics
Ran Ran,
Javier Muñoz Briones,
Smrutiti Jena,
Nicole L. Anderson,
Matthew R. Olson,
Leopold N. Green,
Douglas K. Brubaker
Affiliations
Ran Ran
Center for Global Health and Diseases, Department of Pathology, Case Western Reserve University, Cleveland, OH, USA
Javier Muñoz Briones
Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA; Purdue Interdisciplinary Life Science Program, West Lafayette, IN, USA
Smrutiti Jena
Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA
Nicole L. Anderson
Department of Biological Sciences, Purdue University, West Lafayette, IN, USA
Matthew R. Olson
Department of Biological Sciences, Purdue University, West Lafayette, IN, USA
Leopold N. Green
Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA
Douglas K. Brubaker
Center for Global Health and Diseases, Department of Pathology, Case Western Reserve University, Cleveland, OH, USA; The Blood, Heart, Lung, and Immunology Research Center, Case Western Reserve University, University Hospitals of Cleveland, Cleveland, OH, USA; Corresponding author
Summary: The co-culture of two adult human colorectal cancer cell lines, Caco-2 and HT29, on Transwell is commonly used as an in vitro gut mimic, yet the translatability of insights from such a system to adult human physiological contexts is not fully characterized. Here, we used single-cell RNA sequencing on the co-culture to obtain a detailed survey of cell type heterogeneity in the system and conducted a holistic comparison with human physiology. We identified the intestinal stem cell-, transit amplifying-, enterocyte-, goblet cell-, and enteroendocrine-like cells in the system. In general, the co-culture was fetal intestine-like, with less variety of gene expression compared to the adult human gut. Transporters for major types of nutrients were found in the majority of the enterocytes-like cells in the system. TLR 4 was not expressed in the sample, indicating that the co-culture model is incapable of mimicking the innate immune aspect of the human epithelium.
