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Series GSE80636 Query DataSets for GSE80636
Status Public on Jan 03, 2017
Title Producing enteroendocrine cells from Lgr5+ intestinal stem cells by manipulating quiescence
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Lgr5+ adult intestinal stem cells are highly proliferative throughout life. Single Lgr5+ stem cells can be cultured into 3D epithelial organoids containing all cell types at nearnormal ratios. Culture conditions to generate the main cell types have been established previously, but signals inducing the various types of enteroendocrine cells (EECs) have remained elusive. Here we generate quiescent Lgr5+ stem cells in vitro by inhibition of the EGF-receptor (EGFR) and mitogen-associated protein kinase (MAPK) signaling pathways in organoids, a state that can be readily reversed. Quiescent Lgr5+ stem cells gain a distinct molecular signature, biased towards EEC differentiation. Indeed, combined inhibition of Wnt, Notch and MAPK pathways efficiently generates a diversity of EEC subtypes in vitro. Our observations uncouple Wnt-dependent stem cell maintenance from EGF-dependent proliferation and cell fate choice, and provide an in vitro approach for the study of the elusive EECs.
Overall design We established a stable culture of quiescent Lgr5+ intestinal stem cells in culture. These highly resemble quiescent secretory precursors, which has high EEC differentiation potential. Following on this lead, we elucidated what signals are required to generate EEC cells of all varieties, and provide a method to produce these EEC cells in large numbers.
Contributor(s) Basak O, Beumer J, Wiebrands K, Seno H, van Oudenaarden A, Clevers H
Citation(s) 27939219
Submission date Apr 25, 2016
Last update date May 15, 2019
Contact name Kay Wiebrands
Organization name Hubrecht Institute
Street address Uppsalalaan 8
City Utrecht
ZIP/Postal code 3584CT
Country Netherlands
Platforms (1)
GPL19057 Illumina NextSeq 500 (Mus musculus)
Samples (58)
GSM2132016 Extracted RNA from single cells of EGFR/WNT/Notch inhibited organoids replicate 1
GSM2132017 Extracted RNA from single cells of EGFR/WNT/Notch inhibited organoids replicate 2
GSM2132018 Extracted RNA from single cells of EGFR/WNT/Notch inhibited organoids replicate 3
BioProject PRJNA319552
SRA SRP073803

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Supplementary file Size Download File type/resource
GSE80636_Bulk1.coutc.csv.gz 253.1 Kb (ftp)(http) CSV
GSE80636_Bulk2.coutc.csv.gz 109.7 Kb (ftp)(http) CSV
GSE80636_RAW.tar 6.5 Mb (http)(custom) TAR (of CSV)
GSE80636_cel-seq_barcodes.csv.gz 424 b (ftp)(http) CSV
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Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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