Autophagy contributes to homeostasis in esophageal epithelium where high autophagic vesicle content marks basal cells with limited proliferation and enchanced self-renewal potential [scRNA-Seq]

We used scRNA-seq to examine the cellular and molecular alterations between 3D organoids generated from esophageal epithelial cells with low and high autophagic vesicle content. Esophageal epithelial cells were isolated from C57B6 animals, stained for integrin-b1, DAPI and Cyto-ID amd sorted into two populations: (1) Integrin-b1 positive, DAPI negative, Cyto-ID Low and (2) Integrin-b1 positive, DAPI negative, Cyto-ID High. Then, 500-1000 of sorted cells were seeded per well in 50 μl Matrigel and cultured with mouse organoids media for 15 days. Single cells were isolated from organoids and analyzed via scRNA-Seq.