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

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.

authors

Hahn S,Kim MS,Choi SY,Jeong S,Jee J,Kim HK,Jeong SY,Shin H,Kim HS,Park JS,Yoo J

doi

10.1016/j.bbrc.2018.11.003

subject

Has Abstract

pub_date

2019-01-08 00:00:00

pages

430-439

issue

2

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(18)32401-X

journal_volume

508

pub_type

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