Document Detail


Conditionally immortalized colonic epithelial cell line from a Ptk6 null mouse that polarizes and differentiates in vitro.
MedLine Citation:
PMID:  18205771     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND AND AIMS: PTK6 is an intracellular src-related tyrosine kinase that regulates differentiation in the intestine, where knockout animals have increased proliferative activity and growth characteristics. To explore the phenotype further we attempted to establish epithelial cell lines from the intestinal mucosa.
METHOD: We mated Ptk6 null mice with a tsSV40 large T transgenic mouse (Immortomouse) to obtain null mice carrying the SV40 gene. Intestinal tissues from these mice were cultured.
RESULTS: We established a Ptk6 null epithelial cell line from the colonic mucosa. Consistent with a role of Ptk6 in cell differentiation, these cells have the characteristics of a stable progenitor cell. In monolayer culture, the cells form domes in the monolayer when confluent. When cultured on Transwell filters, the cells polarize and form an electrically resistant barrier. Formation of tight junctions was confirmed by demonstrating expression of ZO1 and occludin at the apical junctions, whereas E-cadherin localized to the basolateral membrane. When cultured in collagen gel, the Ptk6 null cells form complex organoids, some of which resemble cups of cells. These organoids contain cells with differentiated phenotypes. Using immunohistochemistry and confocal microscopy we have been able to identify villin-positive (absorptive cells) and a small percentage of mucin-containing cells (goblet cells) and chromogranin A-positive cells (endocrine cells).
CONCLUSION: This conditionally immortalized cell line represents an excellent cell culture model system for exploring the mechanisms of cell function and epithelial differentiation in the colonic mucosa.
Authors:
Robert H Whitehead; Pamela S Robinson; Janice A Williams; Wenjun Bie; Angela L Tyner; Jeffrey L Franklin
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2008-01-19
Journal Detail:
Title:  Journal of gastroenterology and hepatology     Volume:  23     ISSN:  1440-1746     ISO Abbreviation:  J. Gastroenterol. Hepatol.     Publication Date:  2008 Jul 
Date Detail:
Created Date:  2008-08-14     Completed Date:  2008-10-30     Revised Date:  2014-09-14    
Medline Journal Info:
Nlm Unique ID:  8607909     Medline TA:  J Gastroenterol Hepatol     Country:  Australia    
Other Details:
Languages:  eng     Pagination:  1119-24     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cadherins / metabolism
Cell Differentiation*
Cell Line
Cell Polarity*
Cell Proliferation
Cell Shape
Chromogranin A / metabolism
Colon / cytology,  enzymology*
Electric Impedance
Epithelial Cells / enzymology*
Intestinal Mucosa / cytology,  pathology*
Membrane Proteins / metabolism
Mice
Mice, Knockout
Microfilament Proteins / metabolism
Mucins / metabolism
Occludin
Organoids
Phenotype
Phosphoproteins / metabolism
Tight Junctions / metabolism
Time Factors
Zonula Occludens-1 Protein
src-Family Kinases / deficiency,  genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
CA84239/CA/NCI NIH HHS; CA95103/CA/NCI NIH HHS; DK058404/DK/NIDDK NIH HHS; DK44525/DK/NIDDK NIH HHS; R01 DK044525/DK/NIDDK NIH HHS; R01 DK044525-14/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Cadherins; 0/Chromogranin A; 0/Membrane Proteins; 0/Microfilament Proteins; 0/Mucins; 0/Occludin; 0/Ocln protein, mouse; 0/Phosphoproteins; 0/Tjp1 protein, mouse; 0/Zonula Occludens-1 Protein; 0/villin; EC 2.7.1.-/src-related intestinal kinase, mouse; EC 2.7.10.2/src-Family Kinases
Comments/Corrections

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