Prolactin and epidermal growth factor regulation of the proliferation, morphogenesis, and functional differentiation of normal rat mammary epithelial cells in three dimensional primary culture
Prolactin and epidermal growth factor regulation of the proliferation, morphogenesis, and functional differentiation of normal rat mammary epithelial cells in three dimensional primary culture
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DOI：
10.1002/jcp.1041630216
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发表时间：
1995-05
影响因子：
5.6
通讯作者：
K. Darcy;S. Shoemaker;P. Lee;M. Vaughan;J. Black;M. Ip
中科院分区：
文献类型：
--
作者：
K. Darcy;S. Shoemaker;P. Lee;M. Vaughan;J. Black;M. Ip
利用本实验室开发的三维原代培养模型，研究了泌乳素和表皮生长因子（EGF）对正常大鼠乳腺上皮细胞（MEC）发育的上皮细胞特异性影响。非产乳MEC为球形终芽样乳腺上皮类器官（MEO），分离自短毛期雌性大鼠。培养的MEO发育成精细的多小叶和小叶导管肺泡类器官，由细胞学和功能分化的MEC组成。泌乳素（0.01 ~ 10 μg/ml）和EGF （1 ~ 100 ng/ml）均可诱导细胞生长、广泛的肺泡和多小叶分支形态发生以及酪蛋白积累。在没有泌乳素的情况下培养14天的MEO对泌乳素的有丝分裂、形态形成和产乳作用仍然敏感。同样，在没有EGF的情况下培养的细胞对EGF的有丝分裂和产乳作用仍然敏感，但当在21天培养期的第14天添加EGF时，细胞对其形态形成作用的反应较弱。如果在第一周后停止暴露于催乳素，则有丝分裂和产乳效应的程度下降，但形态发生反应没有下降。在第7天去除EGF也降低了有丝分裂反应，但对产乳反应或形态发生反应的大小没有任何影响。这些研究表明，正常MEC的生理相关发育可以在培养中诱导，并且该模型系统可用于研究催乳素和EGF调节乳腺复杂发育途径的机制。©1995 Wiley‐Liss, Inc。
The epithelial cell‐specific effects of prolactin and epidermal growth factor (EGF) on the development of normal rat mammary epithelial cells (MEC) were evaluated using a three dimensional primary culture model developed in our laboratory. Non‐milk‐producing MEC were isolated as spherical end bud‐like mammary epithelial organoids (MEO) from pubescent virgin female rats. The cultured MEO developed into elaborate multilobular and lobuloductal alveolar organoids composed of cytologically and functionally differentiated MEC. Prolactin (0.01–10 μg/ml) and EGF (1–100 ng/ml) were each required for induction of cell growth, extensive alveolar, as well as multilobular branching morphogenesis, and casein accumulation. MEO cultured without prolactin for 14 days remained sensitive to the mitogenic, morphogenic, and lactogenic effects of prolactin upon subsequent exposure. Similarly, cells cultured in the absence of EGF remained sensitive to the mitogenic and lactogenic effects of EGF, but were less responsive to its morphogenic effects when it was added on day 14 of a 21‐day culture period. If exposure to prolactin was terminated after the first week, the magnitude of the mitogenic and lactogenic effects, but not the morphogenic response was decreased. Removal of EGF on day 7 also reduced the mitogenic response, but did not have any effect on the magnitude of the lactogenic or morphogenic responses. These studies demonstrate that physiologically relevant development of normal MEC can be induced in culture and that this model system can be used to study the mechanisms by which prolactin and EGF regulate the complex developmental pathways operative in the mammary gland. © 1995 Wiley‐Liss, Inc.