PMID- 22564827
OWN - NLM
STAT- MEDLINE
DCOM- 20130110
LR  - 20230603
IS  - 1095-8673 (Electronic)
IS  - 0022-4804 (Print)
IS  - 0022-4804 (Linking)
VI  - 178
IP  - 1
DP  - 2012 Nov
TI  - Intestinal crypts reproducibly expand in culture.
PG  - 48-54
LID - S0022-4804(12)00246-6 [pii]
LID - 10.1016/j.jss.2012.03.037 [doi]
AB  - BACKGROUND: In vitro growth techniques for intestinal crypts and single 
      intestinal stem cells have been recently described, but several questions of 
      translational importance remain unaddressed. The purpose of this study was to 
      first, evaluate if intestinal crypts reproducibly expand in vitro; second, 
      determine the impact of age and region of intestine on crypt growth in vitro; and 
      third, determine the effects of cryopreservation on crypt growth in vitro. 
      METHODS AND MATERIALS: Crypts were harvested from 5 cm of proximal, middle, and 
      distal small intestine of C57BL/6J mice aged 4 wk, 6-8 wk, 12-14 wk, and 18-20 wk 
      (n = 4-6 animals) and cultured. For each region, we determined the efficiency of 
      crypts forming enterospheres (day 1) and progressing to enteroids (day 7). 
      Subsequently, enteroids were passaged and cryopreserved to determine if growth 
      was changed by these manipulations. RESULTS: Forty-three to 99% of intestinal 
      crypts formed enterospheres, with higher efficiency in proximal small intestine 
      and in younger mice. Twenty-five to 64% of enterospheres progressed to budding 
      enteroids within 7 d. In vitro expansion was greater in proximal enteroids. This 
      expansion continued in a logarithmic fashion, with  approximately  97% replating efficiency of 
      isolated enteroid crypt buds. Following cryopreservation,  approximately  90% of enteroids 
      recovered normal proliferative capacity. CONCLUSIONS: Intestinal crypt culture is 
      efficient and significantly expands intestinal tissue in a reproducible manner. 
      Regional and age growth differences may reflect distinct stem cell 
      characteristics or differences in support cells. The ability to culture and 
      expand intestinal tissue in vitro provides a potential translational approach 
      toward understanding and treating patients with short bowel syndrome.
CI  - Copyright (c) 2012 Elsevier Inc. All rights reserved.
FAU - Fuller, Megan K
AU  - Fuller MK
AD  - Department of Surgery, University of North Carolina at Chapel Hill, Chapel Hill, 
      North Carolina, USA.
FAU - Faulk, Denver M
AU  - Faulk DM
FAU - Sundaram, Nambirajan
AU  - Sundaram N
FAU - Shroyer, Noah F
AU  - Shroyer NF
FAU - Henning, Susan J
AU  - Henning SJ
FAU - Helmrath, Michael A
AU  - Helmrath MA
LA  - eng
GR  - U01-DK085547/DK/NIDDK NIH HHS/United States
GR  - R01 CA142826/CA/NCI NIH HHS/United States
GR  - P30 DK034987/DK/NIDDK NIH HHS/United States
GR  - U24 DK085532/DK/NIDDK NIH HHS/United States
GR  - U01 DK085532/DK/NIDDK NIH HHS/United States
GR  - R01 DK092306/DK/NIDDK NIH HHS/United States
GR  - R01-DK083325/DK/NIDDK NIH HHS/United States
GR  - R01 DK083325/DK/NIDDK NIH HHS/United States
GR  - U01 DK085547/DK/NIDDK NIH HHS/United States
GR  - R01 DK092456/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 - 20120410
PL  - United States
TA  - J Surg Res
JT  - The Journal of surgical research
JID - 0376340
SB  - IM
MH  - Adult Stem Cells/cytology
MH  - Animals
MH  - Cell Proliferation
MH  - Cryopreservation/methods
MH  - Intestinal Mucosa/*cytology
MH  - Intestine, Small/cytology
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Organ Culture Techniques/*methods/*standards
MH  - Reproducibility of Results
MH  - Spheroids, Cellular/cytology
MH  - Tissue Engineering/*methods/*standards
MH  - Translational Research, Biomedical
PMC - PMC3654833
MID - NIHMS375560
EDAT- 2012/05/09 06:00
MHDA- 2013/01/11 06:00
PMCR- 2013/11/01
CRDT- 2012/05/09 06:00
PHST- 2012/02/01 00:00 [received]
PHST- 2012/03/19 00:00 [revised]
PHST- 2012/03/20 00:00 [accepted]
PHST- 2012/05/09 06:00 [entrez]
PHST- 2012/05/09 06:00 [pubmed]
PHST- 2013/01/11 06:00 [medline]
PHST- 2013/11/01 00:00 [pmc-release]
AID - S0022-4804(12)00246-6 [pii]
AID - 10.1016/j.jss.2012.03.037 [doi]
PST - ppublish
SO  - J Surg Res. 2012 Nov;178(1):48-54. doi: 10.1016/j.jss.2012.03.037. Epub 2012 Apr 
      10.