Titlebook: Intestinal Stem Cells; Methods and Protocol Paloma Ordóñez-Morán Book 2020 Springer Science+Business Media, LLC, part of Springer Nature 20

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Analysis of Aged Dysfunctional Intestinal Stem Cellss that depend on stem cell in the adult. Intestine is such an organ, and upon aging intestinal regeneration is impaired due to decline of intestinal stem cell function. To determine the aging status of intestine and intestinal stem cells, histological analyses; analyses of the level of proliferation

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Autophagy Detection in Intestinal Stem Cellsagy in intestinal stem cells (ISCs) is important for ISC homeostasis. Here we describe the detailed methods for detecting autophagy in ISCs by observing autophagosomes in GFP-LC3 transgenic mice and quantifying the p62 protein levels. We also describe methods for detecting mitophagy in these cells,

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Single-Cell Transcriptional Profiling of the Intestinal Epitheliumution. By profiling the set of expressed transcripts in each cell, single-cell transcriptomics has allowed for the cataloging of the cellular constituents of multiple organs and tissues, both in health and disease. In addition, these technologies have provided mechanistic insights into cellular func

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Large-Scale Production of Recombinant Noggin and R-Spondin1 Proteins Required for the Maintenance ofem cells. Here, we describe a transient gene expression method for the production of these proteins from human embryo kidney 293 (HEK293) cells cultivated in suspension using orbitally shaken bioreactors. Plasmid DNA was delivered into cells using the cationic polymer polyethylenimine (PEI). The 7-d

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In Vivo Human PSC-Derived Intestinal Organoids to Study Stem Cell Maintenancend associated diseases. Herein, we present a method for renal transplantation of HIOs in immunocompromised mice and subsequent analysis to study intestinal epithelial cell proliferation. In addition, we describe how to generate enteroids from transplanted HIOs. The method highlights the specific ste

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