title: Murine Colonic Organoid Culture System and Downstream Assay Applications.
abstract: Colonic organoids, three-dimensional colonic crypts grown in vitro that show realistic microanatomy, have many potential applications for studying physiology, developmental biology, and pathophysiology of intestinal diseases including inflammatory bowel disease and colorectal cancer. Here, we describe detailed protocols for mouse colonic crypt isolation, organoid culture, and downstream applications. Specific culture strategies including growth factor enriched Matrigel and Wnt and R-spondin conditioned media serve as key factors for enhancing the growth and cost efficiency of colonic organoid cultures.
authors.affiliation: Program in Integrative Nutrition and Complex Diseases, Department of Nutrition & Food Science, Texas A&M University, 112 Cater-Mattil, MS 2253, College Station, TX, 77843-2253, USA.
authors.forename: Yang-Yi
authors.initials: YY
authors.lastname: Fan
authors.affiliation: Program in Integrative Nutrition and Complex Diseases, Department of Nutrition & Food Science, Texas A&M University, 112 Cater-Mattil, MS 2253, College Station, TX, 77843-2253, USA.|Center for Translational Environmental Health Research, Texas A&M University, 112 Cater-Mattil, MS 2253, College Station, TX, 77843-2253, USA.
authors.forename: Laurie A
authors.initials: LA
authors.lastname: Davidson
authors.affiliation: Program in Integrative Nutrition and Complex Diseases, Department of Nutrition & Food Science, Texas A&M University, 112 Cater-Mattil, MS 2253, College Station, TX, 77843-2253, USA. r-chapkin@tamu.edu.|Center for Translational Environmental Health Research, Texas A&M University, 112 Cater-Mattil, MS 2253, College Station, TX, 77843-2253, USA. r-chapkin@tamu.edu.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Chapkin
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology* / Q000378:metabolism; D017077:Culture Media, Conditioned; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D051379:Mice; D009940:Organoids / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D036341:Intercellular Signaling Peptides and Proteins
keywords: In vitro primary culture; Niche-independent crypt expansion; Stem cells