Sugiyama N, Tabuchi Y, Horiuchi T, Obinata M, Furusawa M
Molecular Biology Research Laboratory, Daiichi Pharmaceutical Co., Ltd., Tokyo, Japan.
Exp Cell Res. 1993 Dec;209(2):382-7. doi: 10.1006/excr.1993.1324.
We succeeded in establishing two gastric surface mucous cell lines (designated as GSM06 and GSM10), which produce periodic acid-Schiff (PAS)- and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene. At the permissive temperature (33 degrees C), GSM06 and GSM10 cells grew until confluent monolayers were formed and have now been in culture for more than 9 months with regular passaging. Con A-horseradish peroxidase- and PAS-positive staining indicated that these cells retain the characteristics of gastric surface mucous cells. GSM06 cells showed temperature-sensitive growth in culture and expressed SV40 large T-antigen at the permissive temperature but not at a nonpermissive temperature (39 degrees C). In contrast, GSM10 cells showed temperature-insensitive growth and expressed T-antigen at both permissive and nonpermissive temperatures. To our knowledge, this is the first report of the establishment of gastric surface mucous cell lines from animals. These immortalized cell lines with normal characteristics may serve as good experimental models for the basic and applied biology of gastric surface mucous cells.
我们成功地从携带温度敏感型猿猴病毒40大T抗原基因的成年转基因小鼠胃底黏膜细胞的原代培养物中建立了两种胃表面黏液细胞系（命名为GSM06和GSM10），它们能产生过碘酸希夫（PAS）和刀豆球蛋白A（Con A）阳性糖蛋白。在允许温度（33℃）下，GSM06和GSM10细胞生长直至形成汇合的单层，目前已通过定期传代培养了9个多月。Con A-辣根过氧化物酶和PAS阳性染色表明这些细胞保留了胃表面黏液细胞的特征。GSM06细胞在培养中表现出温度敏感生长，在允许温度下表达SV40大T抗原，但在非允许温度（39℃）下不表达。相比之下，GSM10细胞表现出温度不敏感生长，在允许温度和非允许温度下均表达T抗原。据我们所知，这是首次从动物中建立胃表面黏液细胞系的报道。这些具有正常特征的永生化细胞系可作为胃表面黏液细胞基础生物学和应用生物学的良好实验模型。
