PMID- 22333952
OWN - NLM
STAT- MEDLINE
DCOM- 20120618
LR  - 20220311
IS  - 1528-0012 (Electronic)
IS  - 0016-5085 (Print)
IS  - 0016-5085 (Linking)
VI  - 142
IP  - 5
DP  - 2012 May
TI  - Identification of a cKit(+) colonic crypt base secretory cell that supports 
      Lgr5(+) stem cells in mice.
PG  - 1195-1205.e6
LID - 10.1053/j.gastro.2012.02.006 [doi]
AB  - BACKGROUND & AIMS: Paneth cells contribute to the small intestinal niche of 
      Lgr5(+) stem cells. Although the colon also contains Lgr5(+) stem cells, it does 
      not contain Paneth cells. We investigated the existence of colonic Paneth-like 
      cells that have a distinct transcriptional signature and support Lgr5(+) stem 
      cells. METHODS: We used multicolor fluorescence-activated cell sorting to isolate 
      different subregions of colon crypts, based on known markers, from dissociated 
      colonic epithelium of mice. We performed multiplexed single-cell gene expression 
      analysis with quantitative reverse transcriptase polymerase chain reaction 
      followed by hierarchical clustering analysis to characterize distinct cell types. 
      We used immunostaining and fluorescence-activated cell sorting analyses with in 
      vivo administration of a Notch inhibitor and in vitro organoid cultures to 
      characterize different cell types. RESULTS: Multicolor fluorescence-activated 
      cell sorting could isolate distinct regions of colonic crypts. Four major 
      epithelial subtypes or transcriptional states were revealed by gene expression 
      analysis of selected populations of single cells. One of these, the goblet cells, 
      contained a distinct cKit/CD117(+) crypt base subpopulation that expressed Dll1, 
      Dll4, and epidermal growth factor, similar to Paneth cells, which were also 
      marked by cKit. In the colon, cKit(+) goblet cells were interdigitated with 
      Lgr5(+) stem cells. In vivo, this colonic cKit(+) population was regulated by 
      Notch signaling; administration of a gamma-secretase inhibitor to mice increased the 
      number of cKit(+) cells. When isolated from mouse colon, cKit(+) cells promoted 
      formation of organoids from Lgr5(+) stem cells, which expressed Kitl/stem cell 
      factor, the ligand for cKit. When organoids were depleted of cKit(+) cells using 
      a toxin-conjugated antibody, organoid formation decreased. CONCLUSIONS: cKit 
      marks small intestinal Paneth cells and a subset of colonic goblet cells that are 
      regulated by Notch signaling and support Lgr5(+) stem cells.
CI  - Copyright (c) 2012 AGA Institute. Published by Elsevier Inc. All rights reserved.
FAU - Rothenberg, Michael E
AU  - Rothenberg ME
AD  - Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford, 
      California 94305, USA.
FAU - Nusse, Ysbrand
AU  - Nusse Y
FAU - Kalisky, Tomer
AU  - Kalisky T
FAU - Lee, John J
AU  - Lee JJ
FAU - Dalerba, Piero
AU  - Dalerba P
FAU - Scheeren, Ferenc
AU  - Scheeren F
FAU - Lobo, Neethan
AU  - Lobo N
FAU - Kulkarni, Subhash
AU  - Kulkarni S
FAU - Sim, Sopheak
AU  - Sim S
FAU - Qian, Dalong
AU  - Qian D
FAU - Beachy, Philip A
AU  - Beachy PA
FAU - Pasricha, Pankaj J
AU  - Pasricha PJ
FAU - Quake, Stephen R
AU  - Quake SR
FAU - Clarke, Michael F
AU  - Clarke MF
LA  - eng
GR  - R01 DK080920/DK/NIDDK NIH HHS/United States
GR  - T32 DK0070560/DK/NIDDK NIH HHS/United States
GR  - 5P01CA139490-03/CA/NCI NIH HHS/United States
GR  - 5P30DK056339/DK/NIDDK NIH HHS/United States
GR  - R01 CA100225/CA/NCI NIH HHS/United States
GR  - P30 DK056339/DK/NIDDK NIH HHS/United States
GR  - R01 CA104987/CA/NCI NIH HHS/United States
GR  - P01 CA139490/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20120211
PL  - United States
TA  - Gastroenterology
JT  - Gastroenterology
JID - 0374630
RN  - 0 (Antigens, CD)
RN  - 0 (CD66 antigens)
RN  - 0 (Cell Adhesion Molecules)
RN  - 0 (Hyaluronan Receptors)
RN  - 0 (Lgr5 protein, mouse)
RN  - 0 (Receptors, G-Protein-Coupled)
RN  - 0 (Receptors, Notch)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-kit)
SB  - IM
MH  - Animals
MH  - Antigens, CD/analysis
MH  - Cell Adhesion Molecules/analysis
MH  - Cells, Cultured
MH  - Colon/*cytology
MH  - Flow Cytometry
MH  - Gene Expression Profiling
MH  - Goblet Cells/physiology
MH  - Hyaluronan Receptors/analysis
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Paneth Cells/*chemistry/*physiology
MH  - Proto-Oncogene Proteins c-kit/*analysis
MH  - Receptors, G-Protein-Coupled/*analysis
MH  - Receptors, Notch/physiology
MH  - Single-Cell Analysis
MH  - Stem Cells/chemistry/*physiology
PMC - PMC3911891
MID - NIHMS356740
COIS- Conflicts of interest: This author discloses the following: Stephen R. Quake is a 
      founder, consultant, and shareholder of Fluidigm Corporation. The remaining 
      authors disclose no conflicts.
EDAT- 2012/02/16 06:00
MHDA- 2012/06/19 06:00
PMCR- 2014/02/03
CRDT- 2012/02/16 06:00
PHST- 2011/09/16 00:00 [received]
PHST- 2012/02/01 00:00 [revised]
PHST- 2012/02/02 00:00 [accepted]
PHST- 2012/02/16 06:00 [entrez]
PHST- 2012/02/16 06:00 [pubmed]
PHST- 2012/06/19 06:00 [medline]
PHST- 2014/02/03 00:00 [pmc-release]
AID - S0016-5085(12)00171-0 [pii]
AID - 10.1053/j.gastro.2012.02.006 [doi]
PST - ppublish
SO  - Gastroenterology. 2012 May;142(5):1195-1205.e6. doi: 
      10.1053/j.gastro.2012.02.006. Epub 2012 Feb 11.