Comparison of human induced pluripotent stem cell-derived intestinal tissue (xenografts) with normal adult human colon (NanoString GeoMX DSP)

Tissue samples from human induced pluripotent stem cell-derived intestinal tissue (iPSC-derived intestinal progenitors seeded onto collagen hydrogels) which were transplanted subcutaneously in mice. Mice were sacrificed and tissues were collected after 2, 3 or 4 weeks following engraftment. The tissues were sectioned onto slides and then profiled using NanoString GeoMX DSP (cancer transcriptome atlas). Normal human colon tissue was also profiled as a reference. In both cases, slides were stained for nuclei and cytokeratin. ROIs were selected from in vivo tissue to represent the entire tissue present. For colon reference tissue, crypt apices and bases were selected. Automated segmentation of each ROI using cytokeratin status (positive / negative) was also performed. Gene expression of all genes was evaluated to determine the most differentially expressed (for reference data). These genes and other intestinal genes were specifically investigated further on in vivo samples to ascertain areas of similarity to adult colon and furthermore to demonstrate maturation across timepoints.