Characterization and Application of a Gastric Surface Mucous Cell Line GSM06 Established From Temperature-Sensitive Simian Virus 40 Large T-Antigen Transgenic Mice

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Abstract
It has been indicated that transgenic mouse harboring a temperature-sensitive simian virus 40 large T-antigen gene is useful for establishing cell lines from tissues that have proved difficult to culture in vitro. The gastric surface mucous cell line GSM06 was established from a primary culture of gastric fundic mucosal cells of the transgenic mice. GSM06 cells showed temperature-sensitive growth in culture and expressed large T-antigen at a permissive temperature (33 degrees C) but not at a nonpermissive temperature (39 degrees C). At 39 degrees C, the cells produced periodic acid-Schiff positive glycoconjugates that formed a mucous sheet like the gastric surface mucosa in the stomach. Insulin markedly increased the production of glycoconjugates. In addition, proprotein-processing endoprotease furin suppression retarded cell growth, but accelerated cell differentiation. An air-liquid interface promoted the differentiation of GSM06 cells in a reconstruction culture with nitrocellulose membrane and collagen gel. The gastric surface mucous cell line GSM06 with unique characteristics, therefore, should be useful as an in vitro model of the gastric mucosa for physiological and pharmacological investigations. Moreover, experiments using immortalized cells established in vitro and having specific functions may offer an alternative to experiments using living animals and thereby offer a solution to this ethical issue.

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