Single nucleus RNA sequencing of an in vitro primary adult human colonic epithelium at early, mid, and late time points during long-term culture

We developed a new in vitro model of primary adult human colonic epithelium on a transwell that maintains functional barrier for up to 30 days in culture. The epithelium on the 2-dimensional transwell was derived from normal primary adult human colonoid line Colon 88 at passage 9 (3-dimensional colonoids were grown from isolated crypts of the ascending colon; University of Michigan Translational Tissue Modeling Laboratory). To determine cell composition of the model, the epithelium was collected at early (day 7), mid (day 14), and late (day 28) time points, flash frozen, and stored in liquid nitrogen for nuclei isolation followed by single nuclear RNA sequencing. Nuclei from each time point were derived from two transwells, pooled (n=2 per time point).