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  • Title: Differentiation of Clara cell (distal type) antigen in human fetal bronchial epithelial cell line (HFBE).
    Author: Ochiai A.
    Journal: Exp Toxicol Pathol; 1992 Sep; 44(5):223-34. PubMed ID: 1446158.
    Abstract:
    A human fetal bronchial cell line (HFBE) grew in an undifferentiated pattern under conventional culture conditions. Despite a somewhat fibroblastic shape, however, they still maintained an immuno-reactivity to cytokeratin, carcinoembryonic antigen (CEA) and epithelial membrane antigen (EMA). When grown on a collagen gel in a growth hormone-supplemented medium, their spindle shape became more conspicuous. With the additional supplement of 6 micrograms/ml vitamin A, most of the cells underwent differentiation by producing many bright inclusion bodies which proved to be strongly positive to periodic acid Schiff (PAS) and barely positive to alcian-blue (AB) staining. Electron microscopy revealed well-developed rough endoplasmic reticulum (rER), 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 products containing glycoconjugates of two different molecular weights. The higher molecular weight-product was identified as hyaluronic acid and the lower molecular weight-product as a mixture of neutral glycoproteins containing mainly N-linked glycoproteins whose glycans were of a complex type. While the secretion of hyaluronic acid was inhibited by vitamin A in a dose-dependent manner, that of the neutral glycoproteins was most enhanced by vitamin A in the range from the physiological concentration of 600 ng/ml to 6 micrograms/ml. A monoclonal antibody (SEC-41) generated against the secretory products with the lower molecular weight detected a glycoprotein of approximately 52 kd in the spent culture medium of differentiated HFBE cells. This antibody also reacted with the intra-cytoplasmic secretory granules in these cells. When tested on freeze-sectioned lung tissue, immunohistochemical reactivity of SEC-41 antibody was confined to Clara cells, some type II pneumocytes in the adult lung and respiratory epithelial cells of the fetal lung tissue. Moreover, this antibody could detect the secretory glycoproteins in the broncho-alveolar lavages (BAL) of two human cases. This paper has clearly demonstrated that cells derived from human fetal bronchial epithelium can be cultivated in an undifferentiated precursor state and under appropriate culture conditions they can be stimulated to undergo differentiation into a Clara cell type.
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