Secretory differentiation and cell type identification of a human fetal bronchial epithelial cell line (HFBE).
A human fetal bronchial epithelial cell line (HFBE) grew in an undifferentiated pattern under conventional culture conditions. Despite a somewhat fibroblastic shape the cells maintained immunoreactivity to cytokeratin, carcinoembryonic antigen and epithelial membrane antigen. When grown on a collagen gel in a growth-hormone-supplemented medium, their spindle shape became more conspicuous. With an additional supplement of vitamin A (6 micrograms/ml), most of the cells underwent differentiation by producing many bright inclusion bodies which proved to be strongly positive with periodic acid-Schiff and weakly positive with alcian blue staining. Electron microscopy revealed a well-developed rough endoplasmic reticulum, an enlarged Golgi apparatus and many highly electron-dense secretory granules resembling those of Clara cells. Biochemical analysis demonstrated that HFBE cells cultured on collagen gel with vitamin A secreted hyaluronic acid and neutral glycoproteins containing mainly N-linked glycoproteins whose glycans were of a complex type. A monoclonal antibody (SEC-41) generated against the neutral glycoproteins detected a glycoprotein of approximately 52 kDa in the spent culture medium of differentiated HFBE cells. This antibody also reacted with the intracytoplasmic secretory granules in these cells. When tested on frozen sections of lung tissue, the immunohistochemical reactivity of the SEC-41 antibody was confined to Clara cells, some type II pneumocytes in the adult lung, and respiratory epithelial cells in the fetal lung. Moreover, this antibody could detect secretory glycoprotein in broncho-alveolar lavages from two patients. This paper clearly demonstrates that cells derived from human fetal bronchial epithelium can be cultivated in an undifferentiated precursor state and, under appropriate culture conditions, can be stimulated to undergo differentiation into a Clara cell type.
Ochiai
A
A
Institut für Experimentelle Pathologie, Medizinische Hochschule Hannover, Federal Republic of Germany.
Emura
M
M
Riebe-Imre
M
M
Mohr
U
U
Hilfrich
J
J
Tahara
E
E
Dungworth
D L
DL
D001980:Bronchi / Q000737:chemistry / Q000196:embryology*; D002454:Cell Differentiation; D002460:Cell Line / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D003594:Cytoplasmic Granules / Q000737:chemistry; D004848:Epithelium / Q000196:embryology; D005333:Fetus / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast
D003470:Culture Media