RNA sequencing of chronically damage human colon organoids via Air-liquid interface submergence

We report that repeated rounds of media submergence damage of human colon organoids led to the inability of cells to regrow and respond appropriately to TLR stimulation. We also identified mRNA expression and DNA methylation changes in genes associated with IBD and colon cancer. Overall design: Analysis of two separate human colon organoids from ascending, and transverse colon that have been grown for 10 days, undergone differentiation via air-liquid interface, one round of submergence damage with or without bacterial flagellin stimulation, and five rounds of submergence damage with or without bactieral flagellin stimulation.