Sommerhoff C P, Finkbeiner W E
Department of Pathology, University of California, San Francisco 94143-0506.
Am J Respir Cell Mol Biol. 1990 Jan;2(1):41-50. doi: 10.1165/ajrcmb/2.1.41.
Cellular mechanisms regulating airway secretion, secretory products of individual airway cell types, and control of airway cell growth and differentiation are poorly understood. In order to aid studies of these questions, we have established a system for culturing human tracheobronchial submucosal gland cells. Gland acini were isolated by enzymatic disaggregation from submucosal tissue obtained postmortem from patients without pulmonary diseases and from patients with cystic fibrosis. In culture, acini attached to a collagen substratum, and gland cells proliferated and formed confluent monolayers which were homogeneous by phase microscopy. In contrast to cells of freshly disaggregated acini which expressed either serous or mucous gland cell secretory antigens, in culture virtually all cells (greater than or equal to 95%) concurrently expressed both antigens as assessed by immunocytochemical staining with serous and mucous cell-specific antibodies. Similarly, electron microscopy revealed cells with serous- or mucous-type secretory granules, and cells containing both types of granules. Cultures incorporated 35S into high (greater than 10(6) D) and lower (greater than 700 kD; 150 kD) molecular weight molecules. Cholinergic and adrenergic agonists increased release of radio-labeled secretions. These findings demonstrate that human tracheal gland cells in culture retain immunocytochemical, ultrastructural, and functional features of both differentiated serous and mucous gland cells. This culture system will be useful for studying the biology and pathology of human tracheobronchial submucosal gland cells.
调节气道分泌的细胞机制、个体气道细胞类型的分泌产物以及气道细胞生长和分化的控制目前还知之甚少。为了有助于对这些问题的研究，我们建立了一种培养人气管支气管黏膜下腺细胞的系统。通过酶解从无肺部疾病患者和囊性纤维化患者死后获得的黏膜下组织中分离出腺泡。在培养过程中，腺泡附着于胶原基质上，腺细胞增殖并形成汇合的单层，通过相差显微镜观察这些单层细胞是均匀一致的。与新鲜解离的腺泡细胞不同，新鲜解离的腺泡细胞要么表达浆液性要么表达黏液性腺细胞分泌抗原，而在培养中，通过用浆液性和黏液性细胞特异性抗体进行免疫细胞化学染色评估，几乎所有细胞（大于或等于95%）同时表达这两种抗原。同样，电子显微镜显示有浆液型或黏液型分泌颗粒以及同时含有这两种颗粒的细胞。培养物将35S掺入高分子量（大于10(6) D）和低分子量（大于700 kD；150 kD）分子中。胆碱能和肾上腺素能激动剂增加放射性标记分泌物的释放。这些发现表明，培养中的人气管腺细胞保留了分化的浆液性和黏液性腺细胞的免疫细胞化学、超微结构和功能特征。这种培养系统将有助于研究人气管支气管黏膜下腺细胞的生物学和病理学。
