PMID- 24292484
OWN - NLM
STAT- MEDLINE
DCOM- 20140227
LR  - 20240610
IS  - 1548-7105 (Electronic)
IS  - 1548-7091 (Print)
IS  - 1548-7091 (Linking)
VI  - 11
IP  - 1
DP  - 2014 Jan
TI  - Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their 
      progeny.
PG  - 106-12
LID - 10.1038/nmeth.2737 [doi]
AB  - Although Lgr5(+) intestinal stem cells have been expanded in vitro as organoids, 
      homogeneous culture of these cells has not been possible thus far. Here we show 
      that two small molecules, CHIR99021 and valproic acid, synergistically maintain 
      self-renewal of mouse Lgr5(+) intestinal stem cells, resulting in nearly 
      homogeneous cultures. The colony-forming efficiency of cells from these cultures 
      is ~100-fold greater than that of cells cultured in the absence of CHIR99021 and 
      valproic acid, and multilineage differentiation ability is preserved. We made use 
      of these homogeneous cultures to identify conditions employing simultaneous 
      modulation of Wnt and Notch signaling to direct lineage differentiation into 
      mature enterocytes, goblet cells and Paneth cells. Expansion in these culture 
      conditions may be feasible for Lgr5(+) cells from the mouse stomach and colon and 
      from the human small intestine. These methods provide new tools for the study and 
      application of multiple intestinal epithelial cell types.
FAU - Yin, Xiaolei
AU  - Yin X
AD  - 1] David H. Koch Institute for Integrative Cancer Research, Massachusetts 
      Institute of Technology (MIT), Cambridge, Massachusetts, USA. [2] Center for 
      Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, 
      Cambridge, Massachusetts, USA. [3] Harvard Medical School, Boston, Massachusetts, 
      USA. [4] Harvard-MIT Division of Health Sciences and Technology, MIT, Cambridge, 
      Massachusetts, USA. [5] Harvard Stem Cell Institute, Cambridge, Massachusetts, 
      USA.
FAU - Farin, Henner F
AU  - Farin HF
AD  - Hubrecht Institute for Developmental Biology and Stem Cell Research and 
      University Medical Centre Utrecht, Utrecht, The Netherlands.
FAU - van Es, Johan H
AU  - van Es JH
AD  - Hubrecht Institute for Developmental Biology and Stem Cell Research and 
      University Medical Centre Utrecht, Utrecht, The Netherlands.
FAU - Clevers, Hans
AU  - Clevers H
AD  - Hubrecht Institute for Developmental Biology and Stem Cell Research and 
      University Medical Centre Utrecht, Utrecht, The Netherlands.
FAU - Langer, Robert
AU  - Langer R
AD  - 1] David H. Koch Institute for Integrative Cancer Research, Massachusetts 
      Institute of Technology (MIT), Cambridge, Massachusetts, USA. [2] Harvard-MIT 
      Division of Health Sciences and Technology, MIT, Cambridge, Massachusetts, USA. 
      [3] Department of Chemical Engineering, MIT, Cambridge, Massachusetts, USA.
FAU - Karp, Jeffrey M
AU  - Karp JM
AD  - 1] Center for Biomedical Engineering, Department of Medicine, Brigham and Women's 
      Hospital, Cambridge, Massachusetts, USA. [2] Harvard Medical School, Boston, 
      Massachusetts, USA. [3] Harvard-MIT Division of Health Sciences and Technology, 
      MIT, Cambridge, Massachusetts, USA. [4] Harvard Stem Cell Institute, Cambridge, 
      Massachusetts, USA.
LA  - eng
SI  - GEO/GSE51539
GR  - P30 CA014051/CA/NCI NIH HHS/United States
GR  - R01 DE013023/DE/NIDCR NIH HHS/United States
GR  - DE013023/DE/NIDCR NIH HHS/United States
GR  - CA014051/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 - 20131201
PL  - United States
TA  - Nat Methods
JT  - Nature methods
JID - 101215604
RN  - 0 (Chir 99021)
RN  - 0 (Lgr5 protein, mouse)
RN  - 0 (Pyridines)
RN  - 0 (Pyrimidines)
RN  - 0 (Receptors, G-Protein-Coupled)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 614OI1Z5WI (Valproic Acid)
SB  - IM
MH  - Animals
MH  - Cell Culture Techniques/*instrumentation/*methods
MH  - Cell Differentiation
MH  - Cells, Cultured
MH  - Chromosomes/ultrastructure
MH  - Colon/*cytology
MH  - Flow Cytometry
MH  - Green Fluorescent Proteins/metabolism
MH  - Heterozygote
MH  - Intestine, Small/*cytology
MH  - Karyotyping
MH  - Mice
MH  - Microscopy, Confocal
MH  - Paneth Cells/cytology
MH  - Pyridines/chemistry
MH  - Pyrimidines/chemistry
MH  - Receptors, G-Protein-Coupled/*metabolism
MH  - Signal Transduction
MH  - Stem Cells/*cytology
MH  - Stomach/cytology
MH  - Valproic Acid/chemistry
PMC - PMC3951815
MID - NIHMS548435
COIS- COMPETING FINANCIAL INTERESTS The authors declare no competing financial 
      interests.
EDAT- 2013/12/03 06:00
MHDA- 2014/02/28 06:00
PMCR- 2015/01/01
CRDT- 2013/12/03 06:00
PHST- 2013/07/15 00:00 [received]
PHST- 2013/10/19 00:00 [accepted]
PHST- 2013/12/03 06:00 [entrez]
PHST- 2013/12/03 06:00 [pubmed]
PHST- 2014/02/28 06:00 [medline]
PHST- 2015/01/01 00:00 [pmc-release]
AID - nmeth.2737 [pii]
AID - 10.1038/nmeth.2737 [doi]
PST - ppublish
SO  - Nat Methods. 2014 Jan;11(1):106-12. doi: 10.1038/nmeth.2737. Epub 2013 Dec 1.