Emery N, Place G A, Dodd S, Lhermitte M, David G, Lamblin G, Perini J M, Page A M, Hall R L, Roussel P
Unité INSERM No. 377, Lille, France.
Am J Respir Cell Mol Biol. 1995 Feb;12(2):130-41. doi: 10.1165/ajrcmb.12.2.7865212.
Human bronchial surface epithelial cells were maintained in secondary culture on a collagen gel substrate in a defined, serum-free medium. These conditions have previously been reported to promote mucous cell differentiation. After 3 wk in culture, approximately 40% of the cells were stained by an antibody directed against human respiratory mucin. Analysis of media from cells cultured in the presence of the radioactive precursors [3H]glucosamine and [35S]sulfate revealed that the cells secreted high molecular weight glycoproteins with properties of typical respiratory mucins. In addition, hyaluronic acid and proteoglycans containing chondroitin sulfate and/or heparan sulfate glycosaminoglycans were identified in cell conditioned media. Finally, Western blot analyses showed that the cells secreted lysozyme and mucous proteinase inhibitor, proteins that are generally considered to be markers for submucosal gland serous cells. These results show that human bronchial cells from the surface epithelium in secondary culture secreted a range of glycoconjugates and proteins that were typical secretory products of both mucous and serous cells.
人支气管表面上皮细胞在特定的无血清培养基中的胶原凝胶底物上进行传代培养。此前有报道称，这些条件可促进黏液细胞分化。培养3周后，约40%的细胞被抗人呼吸道黏蛋白抗体染色。对在放射性前体[3H]葡糖胺和[35S]硫酸盐存在下培养的细胞培养基进行分析发现，这些细胞分泌具有典型呼吸道黏蛋白特性的高分子量糖蛋白。此外，在细胞条件培养基中鉴定出含有硫酸软骨素和/或硫酸乙酰肝素糖胺聚糖的透明质酸和蛋白聚糖。最后，蛋白质免疫印迹分析表明，这些细胞分泌溶菌酶和黏液蛋白酶抑制剂，这些蛋白质通常被认为是黏膜下腺浆液细胞的标志物。这些结果表明，传代培养的人支气管表面上皮细胞分泌了一系列糖结合物和蛋白质，它们是黏液细胞和浆液细胞典型的分泌产物。
