PMID- 19398967
OWN - NLM
STAT- MEDLINE
DCOM- 20090723
LR  - 20220408
IS  - 1546-170X (Electronic)
IS  - 1078-8956 (Print)
IS  - 1078-8956 (Linking)
VI  - 15
IP  - 6
DP  - 2009 Jun
TI  - Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell 
      niche.
PG  - 701-6
LID - 10.1038/nm.1951 [doi]
AB  - The in vitro analysis of intestinal epithelium has been hampered by a lack of 
      suitable culture systems. Here we describe robust long-term methodology for small 
      and large intestinal culture, incorporating an air-liquid interface and 
      underlying stromal elements. These cultures showed prolonged intestinal 
      epithelial expansion as sphere-like organoids with proliferation and multilineage 
      differentiation. The Wnt growth factor family positively regulates proliferation 
      of the intestinal epithelium in vivo. Accordingly, culture growth was inhibited 
      by the Wnt antagonist Dickkopf-1 (Dkk1) and markedly stimulated by a fusion 
      protein between the Wnt agonist R-spondin-1 and immunoglobulin Fc (RSpo1-Fc). 
      Furthermore, treatment with the gamma-secretase inhibitor dibenzazepine and 
      neurogenin-3 overexpression induced goblet cell and enteroendocrine cell 
      differentiation, respectively, consistent with endogenous Notch signaling and 
      lineage plasticity. Epithelial cells derived from both leucine-rich 
      repeat-containing G protein-coupled receptor-5-positive (Lgr5(+)) and B lymphoma 
      moloney murine leukemia virus insertion region homolog-1-positive (Bmi1(+)) 
      lineages, representing putative intestinal stem cell (ISC) populations, were 
      present in vitro and were expanded by treatment with RSpo1-Fc; this increased 
      number of Lgr5(+) cells upon RSpo1-Fc treatment was subsequently confirmed in 
      vivo. Our results indicate successful long-term intestinal culture within a 
      microenvironment accurately recapitulating the Wnt- and Notch-dependent ISC 
      niche.
FAU - Ootani, Akifumi
AU  - Ootani A
AD  - Department of Medicine, Division of Hematology, Stanford University School of 
      Medicine, Stanford, California, USA. aootani@stanford.edu
FAU - Li, Xingnan
AU  - Li X
FAU - Sangiorgi, Eugenio
AU  - Sangiorgi E
FAU - Ho, Quoc T
AU  - Ho QT
FAU - Ueno, Hiroo
AU  - Ueno H
FAU - Toda, Shuji
AU  - Toda S
FAU - Sugihara, Hajime
AU  - Sugihara H
FAU - Fujimoto, Kazuma
AU  - Fujimoto K
FAU - Weissman, Irving L
AU  - Weissman IL
FAU - Capecchi, Mario R
AU  - Capecchi MR
FAU - Kuo, Calvin J
AU  - Kuo CJ
LA  - eng
GR  - R01 DK085720-02/DK/NIDDK NIH HHS/United States
GR  - P30 DK56339/DK/NIDDK NIH HHS/United States
GR  - R01 DK069989-05/DK/NIDDK NIH HHS/United States
GR  - R01 DK085720/DK/NIDDK NIH HHS/United States
GR  - R01 CA86065-06/CA/NCI NIH HHS/United States
GR  - R01 DK069989/DK/NIDDK NIH HHS/United States
GR  - U01 DK085527/DK/NIDDK NIH HHS/United States
GR  - P30 DK056339/DK/NIDDK NIH HHS/United States
GR  - R01 DK069989-01/DK/NIDDK NIH HHS/United States
GR  - R01 CA086065/CA/NCI NIH HHS/United States
GR  - U01 DK085527-02/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20090427
PL  - United States
TA  - Nat Med
JT  - Nature medicine
JID - 9502015
RN  - 0 (Basic Helix-Loop-Helix Transcription Factors)
RN  - 0 (Immunoglobulins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Neurog3 protein, mouse)
RN  - 0 (RSPO1 protein, mouse)
RN  - 0 (Receptors, Notch)
RN  - 0 (Thrombospondins)
RN  - 0 (Wnt Proteins)
SB  - IM
MH  - Aging/physiology
MH  - Animals
MH  - Basic Helix-Loop-Helix Transcription Factors/metabolism
MH  - Epithelial Cells/*metabolism
MH  - Immunoglobulins/immunology
MH  - Intestinal Mucosa/*metabolism
MH  - Intestines/ultrastructure
MH  - Mice
MH  - Microscopy, Electron
MH  - Nerve Tissue Proteins/metabolism
MH  - Receptors, Notch/metabolism
MH  - Signal Transduction
MH  - Stem Cell Niche/*metabolism
MH  - Thrombospondins/immunology/metabolism
MH  - Time Factors
MH  - Tissue Culture Techniques/*methods
MH  - Wnt Proteins/*metabolism
PMC - PMC2919216
MID - NIHMS222418
COIS- COMPETING INTERESTS STATEMENT The authors declare competing financial interests: 
      details accompany the full-text HTML version of the paper at 
      http://www.nature.com/naturemedicine/.
EDAT- 2009/04/29 09:00
MHDA- 2009/07/25 09:00
PMCR- 2010/08/10
CRDT- 2009/04/29 09:00
PHST- 2008/12/08 00:00 [received]
PHST- 2009/03/02 00:00 [accepted]
PHST- 2009/04/29 09:00 [entrez]
PHST- 2009/04/29 09:00 [pubmed]
PHST- 2009/07/25 09:00 [medline]
PHST- 2010/08/10 00:00 [pmc-release]
AID - nm.1951 [pii]
AID - 10.1038/nm.1951 [doi]
PST - ppublish
SO  - Nat Med. 2009 Jun;15(6):701-6. doi: 10.1038/nm.1951. Epub 2009 Apr 27.