Taylor G W, Chopra D P, Mathieu P A
Institute of Chemical Toxicology, Wayne State University, Detroit, Michigan.
Epithelial Cell Biol. 1993 Oct;2(4):163-9.
The respiratory tract contains macromolecules produced by various epithelia including tracheal and bronchial mucosa and submucosal glands. The objectives of this study were to elucidate and compare the growth and secretory profiles of epithelial cell cultures derived from the human tracheal (TC) and bronchial mucosa (BC) and submucosal glands (GC). Most experiments were done on third to fourth passage cultures. Secretory glycoconjugates were characterized by a combination of gel filtration and anion-exchange chromatography after enzymic digestion with hyaluronidase of [3H]glucosamine and [35S]sulphate incorporated glycoconjugates secreted into the culture medium. Intracellular mucin-like glycoproteins were characterized by immunohistochemical staining with a human monoclonal respiratory mucin antibody. Results showed that the three cell types exhibited variable growth rates and secretory profiles. Doubling times of GC, BC and TC were 53, 75 and 80 h respectively. Immunocytochemical staining with the mucin antibody demonstrated positive reaction in GC and BC; TC showed no significant reaction. Mucin-like glycoproteins were detected in the spent media of GC and BC whereas TC, under the same conditions, did not produce any detectable amount of the glycoconjugates. Further, the mucin-like materials produced by GC and BC differed in their relative glycosylation and sulphation levels. The production of mucin was independent of substrate and vitamin A as the cultures were propagated on the plastic surfaces and the culture medium lacked vitamin A.
呼吸道包含由各种上皮细胞产生的大分子物质，这些上皮细胞包括气管和支气管黏膜以及黏膜下腺。本研究的目的是阐明并比较源自人气管（TC）、支气管黏膜（BC）和黏膜下腺（GC）的上皮细胞培养物的生长和分泌特征。大多数实验在传代3至4次的培养物上进行。在用透明质酸酶对分泌到培养基中的掺入[³H]葡糖胺和[³⁵S]硫酸盐的糖缀合物进行酶消化后，通过凝胶过滤和阴离子交换色谱相结合的方法对分泌性糖缀合物进行表征。细胞内黏蛋白样糖蛋白通过用人单克隆呼吸道黏蛋白抗体进行免疫组织化学染色来表征。结果表明，这三种细胞类型表现出不同的生长速率和分泌特征。GC、BC和TC的倍增时间分别为53小时、75小时和80小时。用黏蛋白抗体进行免疫细胞化学染色显示GC和BC呈阳性反应；TC未显示明显反应。在GC和BC的用过的培养基中检测到黏蛋白样糖蛋白，而在相同条件下，TC未产生任何可检测量的糖缀合物。此外，GC和BC产生的黏蛋白样物质在相对糖基化和硫酸化水平上有所不同。黏蛋白的产生与底物和维生素A无关，因为培养物在塑料表面上增殖，且培养基中缺乏维生素A。
