Differentiated human alveolar epithelial cells and reversibility of their phenotype in vitro
Differentiated human alveolar epithelial cells and reversibility of their phenotype in vitro
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DOI：
10.1165/rcmb.2006-0410oc
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发表时间：
2007-06-01
影响因子：
6.4
通讯作者：
Mason, Robert J.
中科院分区：
文献类型：
--
作者：
Wang, Jieru;Edeen, Karen;Mason, Robert J.
分化胎儿II型细胞的培养已经有很多年了。然而，对分化的成人II型细胞的研究是有限的。我们使用一种已发表的方法分离II型细胞，并开发了用于维持分化的成人肺泡上皮细胞的原代培养系统，用于体外研究。人II细胞在Matrigel或Matrigel和鼠尾胶原混合物(顶端Access)上培养，在角质形成细胞生长因子、异丁基甲基黄嘌呤、8-溴环腺苷和地塞米松(KiAD)存在下，通过表面活性蛋白(SP)-A、SP-B、SP-C和脂肪酸合成酶的表达和形态观察，检测分化的II型细胞表型。这些细胞含有板层包涵体，顶端有微绒毛。在这两个系统中，细胞似乎分化良好。在顶端通路系统中，II型细胞分化标志物最初减少，然后在培养6d后恢复。KIAD的加入也促进了脂质的合成。相反，在鼠尾胶原(或组织培养塑料)上培养的II型细胞慢慢失去其板层包裹体和表面活性蛋白的表达，并增加I型细胞标志物的表达。表型的表达受培养条件的调节，在体外部分是可逆的。
Cultures of differentiating fetal human type II cells have been available for many years. However, studies with differentiated adult human type II cells are limited. We used a published method for type II cell isolation and developed primary culture systems for maintenance of differentiated adult human alveolar epithelial cells for in vitro studies. Human type II cells cultured on Matrigel (basolateral access) or a mixture of Matrigel and rat tail Collagen (apical access) in the presence of keratinocyte growth factor, isobutylmethylxanthine, 8-bromo-cyclicAMP, and dexamethasone (KIAD) expressed the differentiated type II cell phenotype as measured by the expression of surfactant protein (SP)-A, SP-B, SP-C, and fatty acid synthase and their morphologic appearance. These cells contain lamellar inclusion bodies and have apical microvilli. In both systems the cells appear well differentiated. In the apical access system, type II cell differentiation markers initially decreased and then recovered over 6 d in culture. Lipid synthesis was also increased by the addition of KIAD. In contrast, type II cells cultured on rat tail Collagen (or tissue culture plastic) slowly lose their lamellar inclusions and expression of the surfactant proteins and increase the expression of type I cell markers. The expression of the phenotypes is regulated by the culture conditions and is, in part, reversible in vitro.