METHODS FOR CULTIVATION OF KERATINOCYTES WITH AN AIR-LIQUID INTERFACE
METHODS FOR CULTIVATION OF KERATINOCYTES WITH AN AIR-LIQUID INTERFACE
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DOI：
10.1111/1523-1747.ep12540324
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发表时间：
1983-01-01
影响因子：
6.5
通讯作者：
WOODLEY, D
中科院分区：
文献类型：
--
作者：
PRUNIERAS, M;REGNIER, M;WOODLEY, D
在普通培养中，细胞在人造基质上生长并浸入培养基中。在体内，毛囊间表皮细胞生长在基底膜上并暴露于空气中。在使这些细胞的培养更具生理性的第一次努力中，将培养的细胞升高到空气-培养基界面似乎是合理的。表皮细胞可以通过使用保持在刚性支持物上的胶原凝胶来培养。它们也可以生长在覆盖有基底膜等效物（BME）的硝化细胞滤器上，所述基底膜等效物（BME）预先由牛角膜内皮细胞沉积。通过提高培养物的空气介质界面有一些证据表明，一个更完整的分化，作为评估的形态标准。然而，在生物化学上，培养物的升高似乎不诱导那些在浸没培养物中不表达的角蛋白多肽的合成。表皮细胞也可以通过在真皮基质或真皮等同物上培养来培养。当它们在倒置的死猪皮上培养时，表皮细胞合成膜包被颗粒（MCG）。浸泡对照组未发现MCG。通过培养表皮细胞悬液死亡deepidermized真皮（DED），除了角蛋白模式的分化的所有形态学标记。此外，高分子量角蛋白多肽的部分再表达发生。然而，角蛋白由培养细胞的完全表达取决于真皮基质的过滤作用（培养物从下面进料），而不是暴露于空气-液体界面。总之，有几种方法可用于在空气-液体界面培养表皮细胞，这些方法在研究这些细胞对表观遗传影响的反应中是有意义的。
In ordinary cultures, cells are grown on artificial substrates and immersed in culture medium. In vivo, interfollicular epidermal cells grow on the basement membrane and are exposed to air. In a first effort to render the culture of these cells more physiological it seems legitimate to raise the cultured cells to the air-medium interface. Epidermal cells can be raised by the use of collagen gels maintained on a rigid support. They can also be grown on nitrocellouse filters coated with a basement-membrane equivalent (BME) previously deposited by bovine corneal endothelial cells. By raising the cultures to the air-medium interface there is some evidence of a more complete differentiation, as evaluated by morphologic criteria. However, biochemically, the raising of the cultures does not seem to induce the synthesis of those keratin polypeptides which are not expressed in immersed cultures. Epidermal cells can also be raised by culturing them on dermal substrates or dermal equivalents. When they were cultured on inverted dead pig skin, epidermal cells synthesized membrane-coating granules (MCG). MCG were not found in immersed controls. By culturing epidermal-cell suspensions on dead deepidermized dermis (DED), all morphologic markers of differentiation were seen except the keratin pattern. In addition, partial reexpression of high-molecular-weight keratin polypeptides occurred. However, the complete expression of keratins by cultured cells depends on the filtering action of the dermal substrate (the cultures are fed from underneath) more than on exposure to the air-liquid interface. In summary, several methods are available to culture epidermal cells at the air-liquid interface that are of interest in an investigation of the response of these cells to epigenetic influences.