Stem Cell

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    From 3D to 2D: Harmonization of Protocols for Two-dimensional Cultures on Cell Culture Inserts of Intestinal Organoids from Various Species

    In the expanding field of intestinal organoid research, various protocols for three- and two-dimensional organoid-derived cell cultures exist. Two-dimensional organoid-derived monolayers are used to overcome some limitations of three-dimensional

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    Fluidigm Based Single-cell Gene Expression Library Preparation from Patient-derived Small Intestinal Organoids
    Authors:  Kohei Suzuki and Ryuichi Okamoto, date: 10/05/2020, view: 2937, Q&A: 0
    In this protocol, we describe our methods to isolate crypts from patients' biopsy samples and to culture human intestinal stem cells as it’s called “organoid.” Beyond that, we describe how to dissociate organoids cells into single cells for ...
    Gene Expression Analysis of Sorted Cells by RNA-seq in Drosophila Intestine
    Authors:  Jun Chen, Jia Li, Huaiwei Huang and Rongwen Xi, date: 12/20/2016, view: 14215, Q&A: 1
    RNA sequencing (RNA-seq) has become a popular method for profiling gene expression. Among many applications, one common purpose is to identify differentially expressed genes and pathways in different biological or pathological conditions. This ...
    Immunofluorescent Staining of Mouse Intestinal Stem Cells
    Authors:  Kevin P. O’Rourke, Lukas E Dow and Scott W Lowe, date: 02/20/2016, view: 31041, Q&A: 1
    Immunofluorescent staining of organoids can be performed to visualize molecular markers of cell behavior. For example, cell proliferation marked by incorporation of nucleotide (EdU), or to observe markers of intestinal differentiation including ...
    Isolation, Culture, and Maintenance of Mouse Intestinal Stem Cells
    Authors:  Kevin P. O’Rourke, Sarah Ackerman, Lukas E Dow and Scott W Lowe, date: 02/20/2016, view: 26522, Q&A: 6
    In this protocol we describe our modifications to a method to isolate, culture and maintain mouse intestinal stem cells as crypt-villus forming organoids. These cells, isolated either from the small or large intestine, maintain self-renewal and ...
    Isolation and Flow-cytometric Analysis of Mouse Intestinal Crypt Cells
    Authors:  Dayanira Alsina-Beauchamp, Paloma del Reino and Ana Cuenda, date: 11/05/2015, view: 14894, Q&A: 0
    The intestinal epithelial layer forms tubular invaginations into the underlying connective tissue of the lamina propria. These structures, termed crypts, are the basic functional unit of the intestine. Colon crypts and the surrounding lamina propria ...
    Immunolabelling of Thin Slices of Mouse Descending Colon and Jejunum
    Authors:  Julien Bellis, Isabelle Duluc, Jean-Noël Freund and Jan R. De Mey, date: 10/20/2013, view: 9099, Q&A: 0
    This protocol describes a method for efficient immunolabelling of thin tissue slices containing a few rows of intact intestinal crypts, which yields large numbers of them being oriented favorably for recording stacks of optical sections aligned with ...



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