Wu R, Martin W R, Robinson C B, St George J A, Plopper C G, Kurland G, Last J A, Cross C E, McDonald R J, Boucher R
California Primate Research Center, University of California, Davis 95616.
Am J Respir Cell Mol Biol. 1990 Nov;3(5):467-78. doi: 10.1165/ajrcmb/3.5.467.
The effects of culture conditions on growth and differentiation of human tracheobronchial epithelial (HTBE) cells have been defined. Epithelial cells were dissociated from tissues by protease treatment and were plated on tissue culture dishes in F12 medium supplemented with insulin, transferrin, epidermal growth factor, hydrocortisone, cholera toxin, bovine hypothalamus extract, and retinol. HTBE cells did not express any mucociliary function (ciliogenesis or mucin secretion) on tissue culture plastic, but they could be passaged 3 to 5 times with a total of 10 to 25 population doublings. Cells from early passages re-express both these functions when transplanted to tracheal grafts. When tissue culture plates were coated with collagen film or collagen gel substrata, cell attachment and proliferation were stimulated. However, the expression of mucous cell function in culture occurred only when cells were plated on collagen gel substrata and vitamin A (retinol) was present in the medium. Mucous cell differentiation under optimal conditions was defined by ultrastructural studies, by immunologic studies with mucin-specific monoclonal antibodies, and by carbohydrate and amino acid compositional analyses of mucin-like glycoproteins purified from culture medium. These results demonstrate for the first time that HTBE cells can express mucin synthesis and secretion under appropriate culture conditions.
已明确培养条件对人气管支气管上皮（HTBE）细胞生长和分化的影响。通过蛋白酶处理将上皮细胞从组织中解离出来，并接种于添加了胰岛素、转铁蛋白、表皮生长因子、氢化可的松、霍乱毒素、牛下丘脑提取物和视黄醇的F12培养基中的组织培养皿上。HTBE细胞在组织培养塑料上不表达任何黏液纤毛功能（纤毛形成或黏蛋白分泌），但它们可以传代3至5次，总共经历10至25次群体倍增。早期传代的细胞移植到气管移植物后会重新表达这两种功能。当组织培养板用胶原膜或胶原凝胶基质包被时，细胞黏附和增殖受到刺激。然而，只有当细胞接种于胶原凝胶基质且培养基中存在维生素A（视黄醇）时，培养中的黏液细胞功能才会表达。在最佳条件下的黏液细胞分化通过超微结构研究、用黏蛋白特异性单克隆抗体进行的免疫学研究以及对从培养基中纯化的黏蛋白样糖蛋白的碳水化合物和氨基酸组成分析来确定。这些结果首次证明HTBE细胞在适当的培养条件下可以表达黏蛋白合成和分泌。
