Chopra D P, Shoemaker R L, Taylor G W, Mathieu P A
Institute of Chemical Toxicology, Wayne State University, Detroit, Michigan 48201.
In Vitro Cell Dev Biol. 1991 Jan;27(1):13-20. doi: 10.1007/BF02630889.
Cultures of normal human tracheal gland epithelial cells that exhibit functional differentiation have been propagated in serum-free medium supplemented with insulin (5 micrograms/ml), epidermal growth factor (10 ng/ml), hydrocortisone (0.5 micrograms/ml), and bovine pituitary extract (25 micrograms/ml). The cells retain many characteristics of epithelial cells including microvilli on cell surfaces, desmosomes between cells, and tonofilaments in the cytoplasm. In addition, they exhibit keratin-positive titers and react positively with Peanut agglutinin, which is specific for the disaccharide beta-D-galactose-(1----3)N-acetyl D-galactosamine, a major component of mucin glycoprotein. The cells also exhibit normal Cl- channel activity which was enhanced by the cAMP agonist Forskolin. The major component of the cellular secretion was hyaluronic acid; approximately 10% of the void volume material was resistant to hyaluronidase and may contain material similar to mucin glycoprotein. Some of the cell cultures have been maintained in serum-free conditions for 6 to 7 passages. This model will be important to study regulation of ion-channel activities and mucous glycoprotein secretion and to compare such regulations with the tracheal mucosal epithelial cells already established.
已在补充有胰岛素（5微克/毫升）、表皮生长因子（10纳克/毫升）、氢化可的松（0.5微克/毫升）和牛垂体提取物（25微克/毫升）的无血清培养基中培养出具有功能分化的正常人气管腺上皮细胞。这些细胞保留了上皮细胞的许多特征，包括细胞表面的微绒毛、细胞间的桥粒以及细胞质中的张力丝。此外，它们呈现角蛋白阳性滴度，并与花生凝集素呈阳性反应，花生凝集素对二糖β-D-半乳糖-（1→3）N-乙酰-D-半乳糖胺具有特异性，而二糖是粘蛋白糖蛋白的主要成分。这些细胞还表现出正常的氯离子通道活性，该活性可被环磷酸腺苷激动剂福斯高林增强。细胞分泌物的主要成分是透明质酸；约10%的空体积物质对透明质酸酶有抗性，可能含有类似于粘蛋白糖蛋白的物质。一些细胞培养物已在无血清条件下传代6至7次。该模型对于研究离子通道活性和粘蛋白糖蛋白分泌的调节以及将此类调节与已建立的气管黏膜上皮细胞进行比较具有重要意义。
