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

Authors
 Soojung Hahn  ;  Mi Sun Kim  ;  Seon Young Choi  ;  Sukin Jeong  ;  JooHyun Jee  ;  Han Kyung Kim  ;  Sang Yun Jeong  ;  Hyunsoo Shin  ;  Hyoung Sun Kim  ;  Joon Seong Park  ;  Jongman Yoo 
Citation
 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, Vol.508(2) : 430-439, 2019 
Journal Title
 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 
ISSN
 0006-291X 
Issue Date
2019
MeSH
Adult Stem Cells/cytology ; Adult Stem Cells/drug effects ; Adult Stem Cells/metabolism ; Amides/pharmacology ; Animals ; Cell Self Renewal/drug effects ; Cell Self Renewal/genetics ; Cell Self Renewal/physiology ; Culture Media, Serum-Free ; Flavonoids/pharmacology ; Gene Expression/drug effects ; Humans ; Intestinal Mucosa/cytology ; Intestinal Mucosa/metabolism ; Mice ; Mice, Transgenic ; Organoids/cytology ; Organoids/growth & development ; Organoids/metabolism* ; Pyrazoles/pharmacology ; Pyridines/pharmacology ; Pyrimidines/pharmacology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Receptors, G-Protein-Coupled/genetics ; Receptors, G-Protein-Coupled/metabolism* ; Thiosemicarbazones/pharmacology ; Valproic Acid/pharmacology
Keywords
3D culture ; Lgr5+ stem cell ; Organoid ; Stemness
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.
Full Text
https://www.sciencedirect.com/science/article/pii/S0006291X1832401X
DOI
10.1016/j.bbrc.2018.11.003
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Surgery (외과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Hyung Sun(김형선) ORCID logo https://orcid.org/0000-0002-9002-3569
Park, Joon Seong(박준성) ORCID logo https://orcid.org/0000-0001-8048-9990
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/169978
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