ヒト小唾液腺培養細胞から腺房細胞の樹立

Abstract

A piece of minor salivary gland tissue obtained from the lip of a 16 years old female was minced to about 3 mm3 by fine pincettes and cultured with 10% FCS containing MEM supplemented with EGF (10 ng/ml), fungizon (10 mcg/ml) and kanamycin (60 mcg/ml) in a 5% CO2 incubator. Many small bubbles of saliva were found on the surface of the fragments after 3 to 4 days of incubation and outgrowth of cells from the fragments was observed from 7 days of incubation. Monolayer cells of the outgrowth were trypsinized and passaged with fresh culture medium. At the 8th passage, monolayer cells were infected with SV40 at moi 100PFU/cell. After 18 hour-incubation, the suspension of the infected cells was incubated at densities of 10(4) and 10(3) cells/dish within 0.33% agar containing culture medium. Transformed colonies were picked up from soft agar medium and 3 of the 28 colonies were identified as being acinar cells of the salivary gland, since secretory granules and mucosubstances were specifically proved in the cytoplasm of these cells after 2 to 4 days of incubation. One of the typical clone cells was named HA-16 cells. However, the appearances of the secretory granules and mucins in the cytoplasm of the HA-16 cells depended on the cellular growth cycle, i.e. secretory granules and mucins were not found in the growing cells (G1-S-G2-M phase) but many secretory granules and mucins could be recognized in the non-dividing cells (G0 phase).(ABSTRACT TRUNCATED AT 250 WORDS)