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Leucine-rich repeat-containing G-protein coupled receptor 5 enriched organoids under chemically-defined growth conditions

Author:
Hahn, Soojung, Kim, Mi Sun, Choi, Seon Young, Jeong, Sukin, Jee, JooHyun, Kim, Han Kyung, Jeong, Sang Yun, Shin, Hyunsoo, Kim, Hyoung Sun, Park, Joon Seong, Yoo, Jongman
Source:
Biochemical and biophysical research communications 2019 v.508 no.2 pp. 430-439
ISSN:
0006-291X
Subject:
G-protein coupled receptors, G-proteins, culture media, epidermal growth factor, epithelial cells, epithelium, intestines, mice, stem cells, transgenic animals, valproic acid
Abstract:
An organoid is a complex, multi-cell three-dimensional (3D) structure that contains tissue-specific cells. Epithelial stem cells, which are marked by leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5), have the potential for self-renewal and expansion as organoids. However, in the case of intestinal organoids from Lgr5-EGFP-IRES-CreERT2 transgenic mice, in vitro expansion of the Lgr5 expression is limited in a culture condition supplemented with essential proteins, such as epidermal growth factor (E), noggin (N), and R-spondin 1 (R). In this study, we hypothesized that self-renewal of Lgr5+ stem cells in a 3D culture system can be stimulated by defined compounds (CHIR99021, Valproic acid, Y-27632, and A83-01). Our results demonstrated that dissociated single cells from organoids were organized into a 3D structure in the four compounds containing the ENR culture medium in a 3D and two-dimensional (2D) culture system. Moreover, the Lgr5 expression level of organoids from the ENR- and compound-containing media increased. Furthermore, the conversion of cultured Lgr5+ stem cells from 2D to 3D was confirmed. Therefore, defined compounds promote the expansion of Lgr5+ stem cells in organoids.
Agid:
6225455