Unknown,Transcriptomics,Genomics,Proteomics

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RNA Seq of colonic Lgr5+ epithelial stem cells in Il22+/+ and Il22-/- mice during homeostasis and 24h after irradiation with 8 Gy

ABSTRACT: To investigate the influence of Interleukin-22 (IL-22) on colonic intestinal stem cells, we assessed gene expression in these cells during homeostasis and after induction of DNA damage. IL-22 is a lymphocyte-derived cytokine that targets exclusively non-hematopoietic cells. The receptor is expressed on intestinal epithelial cells, including Lgr5+ stem cells. The colonic Lgr5+ epithelial stem cells were highly purified as DAPI-EpCam+CD45-CD24MedLgr5+. DNA damage was induced by whole body irradiation with 8 Gy and cells were isolated 24h after exposure. The following populations were analyzed: Wildtype, unirradiated (Ctrl) Il22-/-, unirradiated (Ctrl) Wildtype, 24h after 8Gy Il22-/-, 24h after 8Gy

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Mus musculus

SUBMITTER: Pedro Hernandez Cerda

PROVIDER: E-MTAB-6723 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The endodermal lining of the adult gastro-intestinal tract harbors stem cells that are responsible for the day-to-day regeneration of the epithelium. Stem cells residing in the pyloric glands of the stomach and in the small intestinal crypts differ in their differentiation program and in the gene repertoire that they express. Both types of stem cells have been shown to grow from single cells into 3D structures (organoids) in vitro. We show that single adult Lgr5-positive stem cells, isolated from small intestinal organoids, require Cdx2 to maintain their intestinal identity and are converted cell-autonomously into pyloric stem cells in the absence of this transcription factor. Clonal descendants of Cdx2null small intestinal stem cells enter the gastric differentiation program instead of producing intestinal derivatives. Conversely, forced expression of Cdx2 in gastric organoids results in their intestinalization. The intestinal genetic program is thus critically dependent on the single transcription factor encoding gene Cdx2. Small intestinal crypts and stomach glands were isolated from Cdx2-/fl / Lgr5-EGFP-CreERT2 mice and cultured for a week in order to generate small intestinal (SI) and stomach (Sto) in vitro organoids. The Lgr5-CreERT2 enzyme activity has been induced by overnight 4-hydroxytamoxifen induction. Tamoxifen treated and untreated Lgr5-EGFPhi SI and Sto stem cells were FACS sorted and seeded back into ENRWfg (Sto med) culture conditions in order to generate Cdx2-/fl small intestinal (Control SI), Cdx2null small intestinal (Cdx2null SI) and Cdx2-/fl stomach (Control Sto) clonal organoids. Cdx2-/fl SI organoids and Cdx2-/fl Sto organoids have been also cultured in ENR (SI med) to induce differentiation. After some passages of clonal organoid expansion, RNA was isolated from Control SI, Cdx2null SI and Control Sto Lgr5-EGFPhi FACS sorted stem cell populations and from smal intestinal and stomach organoids cultured in different conditions and hybridized on Affymetrix Mouse Gene ST 1.1 arrays.

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RNA Seq of colonic Lgr5+ epithelial stem cells in Il22+/+ and Il22-/- mice during homeostasis and 24h after irradiation with 8 Gy

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