PMID- 24232249
OWN - NLM
STAT- MEDLINE
DCOM- 20140826
LR  - 20220408
IS  - 1750-2799 (Electronic)
IS  - 1754-2189 (Print)
IS  - 1750-2799 (Linking)
VI  - 8
IP  - 12
DP  - 2013 Dec
TI  - In vitro expansion and genetic modification of gastrointestinal stem cells in 
      spheroid culture.
PG  - 2471-82
LID - 10.1038/nprot.2013.153 [doi]
AB  - It is useful to be able to grow enriched populations of stem cells in vitro. 
      Growth of stem cells as tissue spheroids is a key methodology permitting 
      sustainable culture of adult epithelial cells. Gastrointestinal stem cells can be 
      propagated by using conditioned medium from a supportive cell line (L-WRN). This 
      protocol describes how to prepare conditioned medium and how to culture stem 
      cell-enriched epithelial spheroids from the mouse gastrointestine. These 
      spheroids are also amenable to genetic modification with recombinant 
      lentiviruses. This system enables many types of cell biological assays that have 
      been performed with immortalized cell lines to be applied to spheroids. Isolation 
      of epithelial cell units from mice takes up to 2 h, and stem cell-enriched 
      gastrointestinal spheroids are obtained within 3 d. Genetically modified 
      spheroids with lentiviruses can be obtained in 2 weeks.
FAU - Miyoshi, Hiroyuki
AU  - Miyoshi H
AD  - Department of Pathology and Immunology, Washington University School of Medicine, 
      St. Louis, Missouri, USA.
FAU - Stappenbeck, Thaddeus S
AU  - Stappenbeck TS
LA  - eng
GR  - P30 DK052574/DK/NIDDK NIH HHS/United States
GR  - R01 DK071619/DK/NIDDK NIH HHS/United States
GR  - P30-DK52574/DK/NIDDK NIH HHS/United States
GR  - DK071619/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 - 20131114
PL  - England
TA  - Nat Protoc
JT  - Nature protocols
JID - 101284307
SB  - IM
MH  - Animals
MH  - *Cell Culture Techniques
MH  - Cell Line
MH  - Female
MH  - Gastrointestinal Tract/*cytology
MH  - HEK293 Cells
MH  - Humans
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - *Spheroids, Cellular
MH  - Stem Cells/*cytology
PMC - PMC3969856
MID - NIHMS557701
COIS- COMPETING FINANTIAL INTERESTS The authors declare no competing financial 
      interests.
EDAT- 2013/11/16 06:00
MHDA- 2014/08/27 06:00
PMCR- 2014/12/01
CRDT- 2013/11/16 06:00
PHST- 2013/11/16 06:00 [entrez]
PHST- 2013/11/16 06:00 [pubmed]
PHST- 2014/08/27 06:00 [medline]
PHST- 2014/12/01 00:00 [pmc-release]
AID - nprot.2013.153 [pii]
AID - 10.1038/nprot.2013.153 [doi]
PST - ppublish
SO  - Nat Protoc. 2013 Dec;8(12):2471-82. doi: 10.1038/nprot.2013.153. Epub 2013 Nov 
      14.