Single-Cell RNA Sequencing Analysis of Intestinal organoids expanded in either IntestiCult™ Plus or IntestiCult™ OGM (Human)

Conventional Intestinal organoids such as those supported by IntestiCult™ Organoid Growth medium (Human) often fail to recapitulate the cellular diversity of the intestinal epithelium and are typically comprised of Lgr5+ stem cells, transit amplifying cells, but few other cell types. IntestiCult™ Plus Organoid Growth Medium is designed to support both the expansion and differentiation of primary human intestinal organoids. Here we sought to compare the cellular diversity and transcriptomic profiles of primary organoids from 3 intestinal regions of a single donor, expanded with both IntestiCult™ OGM (Human) and IntestiCult™ Plus. Overall design: We expanded duodenal, ileal and colonic organoids derived from a single human donor in either IntestiCult™ OGM (Human) or IntestiCult™ Plus. These organoids had been previously expanded with IntestiCult™ OGM and cryostored in Cryostor10. A single vial of each intestinal region was recovered from cryostorage and split into groups expanded with either IntestiCult™ OGM or IntestiCult™ Plus. These cultures were then passaged on a 7 day schedule three times. At the end of the third passage, The organoids were removed with the Matrigel dome with Gentle cell dissociation reagent and cells were dissociated with 0.05% Trypsin-EDTA for analysis. Single cell capture and library prep was performed using a combination of Illumina Single Cell 3' RNA Prep and 10X Genomics Chromium platforms in order to compare technologies.