Establishment and characterization of novel in vitro models of gastric chief cell and metaplastic lineages

Abstract

Investigations of preneoplastic metaplasias in the stomach are limited by the lack of cell models for mature mucosal cells and metaplasias. We developed in vitro models of chief cells and spasmolytic polypeptide‐expressing metaplasia (SPEM) cells to study the transdifferentiation of chief cells into SPEM. Utilizing the Immortomouse, which is completely transgenic for tempertature‐dependent TAg, chief cells were isolated and cultured from untreated mice. SPEM cells were isolated from mice treated with L635, an acute SPEM induction model. mRNA and protein expression profiles were used to characterize the cultures. ImChief cultures express most chief cell markers with immunostaining for pepsinogen and Rab3D‐containing granules. After 2 weeks at 39°C, ImChief cells cease proliferating and resemble a more differentiated state similar to in vivo chief cells. ImSPEM cells express the SPEM markers TFF2 and HE4 and produce HE4‐containing granules. They lack expression of most other lineage specific markers. ImSPEM cells retain their proliferative ability for at least 1 month at 39°C, resembling the in vivo properties of SPEM cells. Our results indicate that the expression and proliferation profiles of the novel ImChief and ImSPEM cultures resemble in vivo chief and SPEM cell lineages. These cultures provide the first in vitro system for studying the molecular mechanisms of the metaplastic transition in the stomach.Department of Veterans Affairs Merit Review Award and NIH RO1 DK071590