EFFECT OF CELL-SHAPE CHANGE ON THE FUNCTION AND DIFFERENTIATION OF RABBIT MAMMARY CELLS IN CULTURE
EFFECT OF CELL-SHAPE CHANGE ON THE FUNCTION AND DIFFERENTIATION OF RABBIT MAMMARY CELLS IN CULTURE
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DOI：
10.1083/jcb.96.5.1425
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发表时间：
1983-01-01
影响因子：
7.8
通讯作者：
KRAEHENBUHL, JP
中科院分区：
文献类型：
--
作者：
HAEUPTLE, MT;SUARD, YLM;KRAEHENBUHL, JP
细胞形状，细胞分化和组织形貌的诱导和维持功能分化的作用进行了研究，在兔乳腺细胞生长的原代培养物上的2维胶原蛋白表面或3维胶原蛋白基质。将中孕兔乳腺分离成单细胞，并通过等密度离心富集上皮细胞。将在旋转振荡器上形成的上皮细胞的小球体（约50个细胞）铺在胶原凝胶上或包埋在胶原凝胶中。将细胞在含血清培养基中培养1天，然后在化学成分确定的培养基中培养25天。在一些实验中，凝胶上的上皮单层在第2天或第5天从培养皿中机械地游离。这些凝胶收缩并形成漂浮的胶原蛋白凝胶。在附着的胶原蛋白凝胶上，几天内就形成了单一细胞类型的扁平单层。细胞合成DNA，直到实现融合，但不积累乳蛋白。催乳素（PRL）未引起形态学变化。在浮动凝胶上，在没有细胞增殖的情况下出现2种细胞类型。与培养基直接接触的细胞变成立方形，并形成典型的分泌细胞的细胞器。PRL诱导的脂肪生成，导致大脂肪滴填充顶端细胞质，酪蛋白和α-乳清蛋白积聚在脂肪滴周围的囊泡中。转铁蛋白检测在存在或不存在的PRL细胞内的小泡，但也在与细胞层接触的胶原蛋白基质。第二类细胞位于分泌细胞层和胶原基质之间，富含微丝，类似肌上皮细胞。在包埋凝胶中，细胞形成中空的导管状结构，其尺寸不断增大。分泌细胞形成典型的管腔，管腔内分泌物增多。在与胶原蛋白凝胶接触时，发现很少有富含微生物活性的细胞。脂肪、酪蛋白和α-的储存和分泌乳白蛋白需要PRL的存在，而转铁蛋白的积累和矢量放电是催乳素独立的。由于DNA的合成和乳蛋白的储存是沿着管状结构随机进行的，因此在顶端和中心之间没有分化梯度。功能极性的建立和细胞分化的诱导可能受到细胞与胶原结构相互作用的性质的影响。形态分化反过来又在脂肪和乳蛋白的合成、储存和分泌中起着重要作用。
The role of cell shape, cytodifferentiation and tissue topography on the induction and maintenance of functional differentiation was examined in rabbit mammary cells grown as primary cultures on 2-dimensional collagen surfaces or in 3-dimensional collagen matrices. Mammary glands from mid-pregnant rabbits were dissociated into single cells, and epithelial cells were enriched by isopycnic centrifugation. Small spheroids of epithelial cells (approximately 50 cells) that formed on a rotary shaker were plated on or embedded in collagen gels. The cells were cultured for 1 d [day] in serum-containing medium and then for up to 25 d in chemically defined medium. In some experiments, epithelial monolayers on gels were mechanically freed from the dishes on day 2 or 5. These gels retracted and formed floating collagen gels. On attached collagen gels, flat monolayers of a single cell type developed within a few days. The cells synthesized DNA until the achievement of confluence but did not accumulate milk proteins. No morphological changes were induced by prolactin (PRL). On floating gels, 2 cell types appeared in the absence of cell proliferation. The cells in direct contact with the medium became cuboidal and developed intracellular organelles typical of secretory cells. PRL-induced lipogenesis, resulting in large fat droplets filling the apical cytoplasm and accumulation of casein and a-lactablumin in vesicles surrounding the fat droplets. Transferrin was detected in the presence or absence of PRL intracellularly in small vesicles but also in the collagen matrix in contact with the cell layer. The 2nd cell type, rich in microfilaments and reminiscent of the myoepithelial cells, was situated between the secretory cell layer and the collagen matrix. In embedding gels, the cells formed hollow ductlike structures, which grew continuously in size. Secretory cells formed typical lumina distended by secretory products. Few microfilament-rich cells were found in contact with the collagen gels. Storage and secretion of fat, caseins and .alpha.-lactalbumin required the presence of PRL, whereas the accumulation and vectorial discharge of transferrin was prolactin independent. There was no differentiation gradient between the tip and the center of the outgrowth, since DNA synthesis and milk protein storage were random along the tubular structures. Establishment of functional polarity and induction of cytodifferentiation may be influenced by the nature of the interaction of the cells with the collagen structure. The morphological differentiation in turn plays an important role in the synthesis, storage and secretion of fat and milk proteins.