Conditionally immortalized colonic epithelial cell line from a Ptk6 null mouse that polarizes and differentiates in vitro.

Journal of Gastroenterology and Hepatology
Robert H WhiteheadJeffrey L Franklin

Abstract

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. 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. 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 ha...Continue Reading

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Citations

Dec 15, 2012·Archives of Microbiology·Angela G LindenstraussRudi F Vogel
Dec 26, 2008·American Journal of Physiology. Gastrointestinal and Liver Physiology·Robert H Whitehead, Pamela S Robinson
Sep 20, 2012·Molecular Cancer Therapeutics·Jessica J GierutAngela L Tyner
Dec 23, 2009·Journal of Cell Science·Helena L Palka-HamblinAngela L Tyner
Dec 24, 2013·Archives of Microbiology·Zhen PengRudi F Vogel
Dec 3, 2014·Biochemical and Biophysical Research Communications·R J HainesM H Wu
Oct 31, 2013·Clinical and Experimental Immunology·M HolzscheiterC Prazeres da Costa
Mar 25, 2017·American Journal of Physiology. Gastrointestinal and Liver Physiology·Tamara StelzlHannelore Daniel
Sep 6, 2014·American Journal of Physiology. Gastrointestinal and Liver Physiology·Victoria G WeisJames R Goldenring

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