Regulation of differentiation and polarized secretion in mammary epithelial cells maintained in culture: extracellular matrix and membrane polarity influences.
Regulation of differentiation and polarized secretion in mammary epithelial cells maintained in culture: extracellular matrix and membrane polarity influences.
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DOI：
10.1083/jcb.105.5.2043
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发表时间：
1987-11
期刊：
影响因子：
--
通讯作者：
Moss L
中科院分区：
文献类型：
--
作者：
Parry G;Cullen B;Kaetzel CS;Kramer R;Moss L
先前的一些研究表明，怀孕小鼠的乳腺上皮细胞只有在培养液中漂浮的基质胶原凝胶中才能保持其分化特征和分泌潜能。通过监测小鼠乳腺上皮细胞系COMMA-1-D的乳蛋白合成和极化分泌，研究了这些培养需求的细胞基础。实验旨在了解细胞-细胞外基质相互作用的可能作用以及满足上皮极性需求的要求。当细胞在漂浮的胶原凝胶上培养时，它们在细胞与基质胶原的界面上聚集了一个由层粘连蛋白、胶原蛋白（IV）和硫酸肝素蛋白聚糖组成的基底层样结构。为了评估这些成分的作用，使用小鼠内胚层细胞系PFHR-9生成了含有这些分子的外源性基底膜。将该基质分离成薄片贴在培养皿上，然后将乳腺细胞镀在上面。结果发现，附着在PFHR-9基质上的培养物表达的β -酪蛋白水平略高于塑料组织培养皿上的细胞，并积累了大量的脂肪滴。然而，β -酪蛋白的水平比漂浮胶原蛋白凝胶培养的低大约四倍。此外，细胞在附着基质上产生的β -酪蛋白不分泌，而是在细胞内迅速降解。然而，如果将附着细胞的PFHR-9基质漂浮在培养基中，β -酪蛋白的表达与漂浮在基质胶原凝胶上培养的细胞相当，酪蛋白也分泌到培养基中。通过在Millicell （Millipore Corp., Bedford， MA）室的硝化纤维素过滤器上培养细胞，测试了培养物漂浮的可能性，以允许进入细胞的基底侧表面。这些腔室允许单层细胞通过基底细胞表面与培养基及其补充的激素和生长因子相互作用。值得注意的是，在这些条件下，α 1-、α 2-和β -酪蛋白的合成与漂浮凝胶和基质上的细胞相当，而且酪蛋白的分泌也很活跃。与滤光片是否涂有基质无关，也得到了类似的结果。（摘要删节为400字）
Several previous studies have demonstrated that mammary epithelial cells from pregnant mice retain their differentiated characteristics and their secretory potential in culture only when maintained on stromal collagen gels floated in the culture medium. The cellular basis for these culture requirements was investigated by the monitoring of milk protein synthesis and polarized secretion from the mouse mammary epithelial cell line, COMMA-1-D. Experiments were directed towards gaining an understanding of the possible roles of cell-extracellular matrix interactions and the requirements for meeting polarity needs of the epithelium. When cells are cultured on floating collagen gels they assemble a basal lamina-like structure composed of laminin, collagen (IV), and heparan sulfate proteoglycan at the interface of the cells with the stromal collagen. To assess the role of these components, an exogenous basement membrane containing these molecules was generated using the mouse endodermal cell line, PFHR-9. This matrix was isolated as a thin sheet attached to the culture dish, and mammary cells were then plated onto it. It was found that cultures on attached PFHR-9 matrices expressed slightly higher levels of beta-casein than did cells on plastic tissue culture dishes, and also accumulated a large number of fat droplets. However, the level of beta-casein was approximately fourfold lower than that in cultures on floating collagen gels. Moreover, the beta-casein made in cells on attached matrices was not secreted but was instead rapidly degraded intracellularly. If, however, the PFHR-9 matrices with attached cells were floated in the culture medium, beta-casein expression became equivalent to that in cells cultured on floating stromal collagen gels, and the casein was also secreted into the medium. The possibility that floatation of the cultures was necessary to allow access to the basolateral surface of cells was tested by culturing cells on nitrocellulose filters in Millicell (Millipore Corp., Bedford, MA) chambers. These chambers permit the monolayers to interact with the medium and its complement of hormones and growth factors through the basal cell surface. Significantly, under these conditions alpha 1-, alpha 2-, and beta- casein synthesis was equivalent to that in cells on floating gels and matrices, and, additionally, the caseins were actively secreted. Similar results were obtained independently of whether or not the filters were coated with matrices.(ABSTRACT TRUNCATED AT 400 WORDS)