PMID- 23840480
OWN - NLM
STAT- MEDLINE
DCOM- 20140121
LR  - 20220430
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 6
DP  - 2013
TI  - Cell lineage identification and stem cell culture in a porcine model for the 
      study of intestinal epithelial regeneration.
PG  - e66465
LID - 10.1371/journal.pone.0066465 [doi]
LID - e66465
AB  - Significant advances in intestinal stem cell biology have been made in murine 
      models; however, anatomical and physiological differences between mice and humans 
      limit mice as a translational model for stem cell based research. The pig has 
      been an effective translational model, and represents a candidate species to 
      study intestinal epithelial stem cell (IESC) driven regeneration. The lack of 
      validated reagents and epithelial culture methods is an obstacle to investigating 
      IESC driven regeneration in a pig model. In this study, antibodies against 
      Epithelial Adhesion Molecule 1 (EpCAM) and Villin marked cells of epithelial 
      origin. Antibodies against Proliferative Cell Nuclear Antigen (PCNA), 
      Minichromosome Maintenance Complex 2 (MCM2), Bromodeoxyuridine (BrdU) and 
      phosphorylated Histone H3 (pH3) distinguished proliferating cells at various 
      stages of the cell cycle. SOX9, localized to the stem/progenitor cells zone, 
      while HOPX was restricted to the +4/'reserve' stem cell zone. Immunostaining also 
      identified major differentiated lineages. Goblet cells were identified by Mucin 2 
      (MUC2); enteroendocrine cells by Chromogranin A (CGA), Gastrin and Somatostatin; 
      and absorptive enterocytes by carbonic anhydrase II (CAII) and sucrase isomaltase 
      (SIM). Transmission electron microscopy demonstrated morphologic and sub-cellular 
      characteristics of stem cell and differentiated intestinal epithelial cell types. 
      Quantitative PCR gene expression analysis enabled identification of 
      stem/progenitor cells, post mitotic cell lineages, and important growth and 
      differentiation pathways. Additionally, a method for long-term culture of porcine 
      crypts was developed. Biomarker characterization and development of IESC culture 
      in the porcine model represents a foundation for translational studies of 
      IESC-driven regeneration of the intestinal epithelium in physiology and disease.
FAU - Gonzalez, Liara M
AU  - Gonzalez LM
AD  - Center for Comparative Medicine and Translational Research, North Carolina State 
      University, Raleigh, North Carolina, United States of America.
FAU - Williamson, Ian
AU  - Williamson I
FAU - Piedrahita, Jorge A
AU  - Piedrahita JA
FAU - Blikslager, Anthony T
AU  - Blikslager AT
FAU - Magness, Scott T
AU  - Magness ST
LA  - eng
GR  - R03DK089126/DK/NIDDK NIH HHS/United States
GR  - UL1TR000083/TR/NCATS NIH HHS/United States
GR  - R01 DK091427/DK/NIDDK NIH HHS/United States
GR  - T32 RR024394/RR/NCRR NIH HHS/United States
GR  - P30 DK034987/DK/NIDDK NIH HHS/United States
GR  - T32RR024394/RR/NCRR NIH HHS/United States
GR  - UL1 TR000083/TR/NCATS NIH HHS/United States
GR  - R03 DK089126/DK/NIDDK NIH HHS/United States
GR  - R01DK091427/DK/NIDDK NIH HHS/United States
GR  - T32 OD011130/OD/NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20130628
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Biomarkers)
SB  - IM
MH  - Animals
MH  - Apoptosis
MH  - Biomarkers/metabolism
MH  - Cell Culture Techniques
MH  - Cell Cycle
MH  - Cell Differentiation
MH  - *Cell Lineage
MH  - Cell Proliferation
MH  - Cells, Cultured
MH  - Epithelial Cells/physiology/ultrastructure
MH  - Goblet Cells/*physiology/ultrastructure
MH  - Intestinal Mucosa/physiology/ultrastructure
MH  - *Regeneration
MH  - Stem Cells/*physiology
MH  - Sus scrofa
PMC - PMC3696067
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2013/07/11 06:00
MHDA- 2014/01/22 06:00
PMCR- 2013/06/28
CRDT- 2013/07/11 06:00
PHST- 2013/04/13 00:00 [received]
PHST- 2013/05/05 00:00 [accepted]
PHST- 2013/07/11 06:00 [entrez]
PHST- 2013/07/11 06:00 [pubmed]
PHST- 2014/01/22 06:00 [medline]
PHST- 2013/06/28 00:00 [pmc-release]
AID - PONE-D-13-15279 [pii]
AID - 10.1371/journal.pone.0066465 [doi]
PST - epublish
SO  - PLoS One. 2013 Jun 28;8(6):e66465. doi: 10.1371/journal.pone.0066465. Print 2013.