Literature DB >> 33659432

Fluidigm Based Single-cell Gene Expression Library Preparation from Patient-derived Small Intestinal Organoids.

Kohei Suzuki1, Ryuichi Okamoto2,3.   

Abstract

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 single-cell analysis as a further application. This protocol should provide investigators sufficient tools to generate human organoids from biopsy samples and to accomplish a stable in-vitro assay system.
Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.

Entities:  

Keywords:  Endoscopic biopsy; Intestinal stem cell; Multiplex PCR; Organoid; Single-cell analysis

Year:  2020        PMID: 33659432      PMCID: PMC7842824          DOI: 10.21769/BioProtoc.3775

Source DB:  PubMed          Journal:  Bio Protoc        ISSN: 2331-8325


  14 in total

1.  The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers.

Authors:  Javier Muñoz; Daniel E Stange; Arnout G Schepers; Marc van de Wetering; Bon-Kyoung Koo; Shalev Itzkovitz; Richard Volckmann; Kevin S Kung; Jan Koster; Sorina Radulescu; Kevin Myant; Rogier Versteeg; Owen J Sansom; Johan H van Es; Nick Barker; Alexander van Oudenaarden; Shabaz Mohammed; Albert J R Heck; Hans Clevers
Journal:  EMBO J       Date:  2012-06-12       Impact factor: 11.598

2.  Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche.

Authors:  Toshiro Sato; Robert G Vries; Hugo J Snippert; Marc van de Wetering; Nick Barker; Daniel E Stange; Johan H van Es; Arie Abo; Pekka Kujala; Peter J Peters; Hans Clevers
Journal:  Nature       Date:  2009-03-29       Impact factor: 49.962

Review 3.  The intestinal crypt, a prototype stem cell compartment.

Authors:  Hans Clevers
Journal:  Cell       Date:  2013-07-18       Impact factor: 41.582

Review 4.  Role of epithelial cells in the pathogenesis and treatment of inflammatory bowel disease.

Authors:  Ryuichi Okamoto; Mamoru Watanabe
Journal:  J Gastroenterol       Date:  2015-07-03       Impact factor: 7.527

5.  Adult stem cells in the small intestine are intrinsically programmed with their location-specific function.

Authors:  Sabine Middendorp; Kerstin Schneeberger; Caroline L Wiegerinck; Michal Mokry; Ronald D L Akkerman; Simone van Wijngaarden; Hans Clevers; Edward E S Nieuwenhuis
Journal:  Stem Cells       Date:  2014-05       Impact factor: 6.277

Review 6.  Integrative single-cell analysis.

Authors:  Tim Stuart; Rahul Satija
Journal:  Nat Rev Genet       Date:  2019-05       Impact factor: 53.242

Review 7.  Human intestinal epithelium in a dish: Current models for research into gastrointestinal pathophysiology.

Authors:  Manuel Noben; Wiebe Vanhove; Kaline Arnauts; Anabela Santo Ramalho; Gert Van Assche; Séverine Vermeire; Catherine Verfaillie; Marc Ferrante
Journal:  United European Gastroenterol J       Date:  2017-07-21       Impact factor: 4.623

Review 8.  Defining a stem cell hierarchy in the intestine: markers, caveats and controversies.

Authors:  Nicholas R Smith; Alexandra C Gallagher; Melissa H Wong
Journal:  J Physiol       Date:  2016-04-24       Impact factor: 6.228

9.  Single-cell analysis of proxy reporter allele-marked epithelial cells establishes intestinal stem cell hierarchy.

Authors:  Ning Li; Maryam Yousefi; Angela Nakauka-Ddamba; Rajan Jain; John Tobias; Jonathan A Epstein; Shane T Jensen; Christopher J Lengner
Journal:  Stem Cell Reports       Date:  2014-10-23       Impact factor: 7.765

10.  Single cell analysis of Crohn's disease patient-derived small intestinal organoids reveals disease activity-dependent modification of stem cell properties.

Authors:  Kohei Suzuki; Tatsuro Murano; Hiromichi Shimizu; Go Ito; Toru Nakata; Satoru Fujii; Fumiaki Ishibashi; Ami Kawamoto; Sho Anzai; Reiko Kuno; Konomi Kuwabara; Junichi Takahashi; Minami Hama; Sayaka Nagata; Yui Hiraguri; Kento Takenaka; Shiro Yui; Kiichiro Tsuchiya; Tetsuya Nakamura; Kazuo Ohtsuka; Mamoru Watanabe; Ryuichi Okamoto
Journal:  J Gastroenterol       Date:  2018-01-27       Impact factor: 7.527

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