Kumar R K, Maronese S E, O'Grady R
School of Pathology, University of New South Wales, Sydney, Australia.
Exp Lung Res. 1997 Sep-Oct;23(5):427-40. doi: 10.3109/01902149709039236.
Numerous investigators have described maintenance of airway epithelial cells from various species in a differentiated state in primary culture. Because the number of cells that can be isolated from the mouse trachea is very small, published techniques are unsuitable for this species. To examine the production of growth factors by murine airway epithelial cells, the authors developed a method for culture of mouse tracheal epithelial cells from explants, in which the population of cells was expanded in the presence of epidermal growth factor and insulin-like growth factor-I, which exhibited synergistic mitogenic activity. After subculture, an essentially pure population of epithelial cells was recovered, with a yield approximately tenfold greater than reported using protease dissociation of cells from the trachea. Culture of the cells at passage 2 on a collagen gel substratum induced differentiation toward a synthetic/secretory phenotype, accompanied by marked diminution in spontaneous and mitogen-induced DNA synthesis without loss of viability. In parallel, secretion of immunoreactive transforming growth factor-beta by the epithelial cells was strikingly increased, but could be partially down-regulated in the presence of mitogenic growth factors.
许多研究人员描述了在原代培养中将来自各种物种的气道上皮细胞维持在分化状态。由于从小鼠气管中分离出的细胞数量非常少，已发表的技术不适用于该物种。为了研究小鼠气道上皮细胞生长因子的产生，作者开发了一种从外植体培养小鼠气管上皮细胞的方法，其中细胞群体在表皮生长因子和胰岛素样生长因子-I存在的情况下得以扩增，这两种因子表现出协同促有丝分裂活性。传代培养后，获得了基本纯的上皮细胞群体，产量比报道的使用蛋白酶从气管中解离细胞的方法高出约十倍。将第2代细胞培养在胶原凝胶基质上可诱导其向合成/分泌表型分化，同时自发和有丝分裂原诱导的DNA合成显著减少，但细胞活力未丧失。与此同时，上皮细胞免疫反应性转化生长因子-β的分泌显著增加，但在有丝分裂生长因子存在的情况下可部分下调。
