Growth of mouse mammary gland end buds cultured in a collagen gel matrix.
Growth of mouse mammary gland end buds cultured in a collagen gel matrix.
复制标题
在胶原凝胶基质中培养的小鼠乳腺末芽的生长。
DOI：
10.1016/0014-4827(82)90231-2
复制
发表时间：
1982
影响因子：
3.7
通讯作者：
Nandi,S
中科院分区：
文献类型：
--
作者：
Richards,J;Guzman,R;Konrad,M;Yang,J;Nandi,S
终芽是乳腺导管生长的末端结构。它们由未分化的上皮细胞组成，在成年和怀孕女性的激素环境下，产生成熟腺体的导管和肺泡。末端芽对了解腺体的发育生物学有重要意义。它们也可能是许多乳腺癌致癌物的目标，因此，对了解乳腺癌的生物学很感兴趣。建立了一种从4周龄C57BL/Crgl小鼠乳腺胶原酶和透明质酸酶消化中分离终芽的方法。在大鼠尾胶原凝胶基质中培养3周，用含有不同血清、激素和生长促进因子组合的培养基喂养。生长对不同培养基的响应是通过拍摄单个端芽菌落随时间使用暗场照明来测量的。使用计算机辅助光密度测定法测定菌落质量，定量测定单个菌落的生长。在添加50%马血清和霍乱毒素或添加较少的血清（15%）和表皮生长因子（EGF）、激素和霍乱毒素的培养基中，末芽表现出最大的生长反应（3周内增长20倍以上，至少48 h翻倍）。最初，菌落呈对数增长，然后随着菌落规模的增加而放缓。这在一定程度上是由于中心细胞停止生长。这种终芽培养系统与分离的小鼠乳腺细胞的培养系统在许多方面有所不同。
End buds are the growing terminal structures of the ducts in the mammary gland. They are made up of undifferentiated epithelial cells that, under the hormonal milieu of adult and pregnant females, give rise to the ducts and alveoli of the mature gland. The end buds are of interest in understanding the developmental biology of the gland. They are also likely targets for many mammary carcinogens and, therefore, of interest in understanding the biology of mammary cancer. A method was developed for isolating end buds as a pure subpopulation from collagenase and hyaluronidase digested 4-week-old C57BL/Crgl mouse mammary glands. They were cultured within a rat tail collagen gel matrix for 3 weeks and fed with media containing various combinations of sera, hormones and growth-promoting factors. Growth in response to the different media was measured by photographing individual end bud colonies over time using dark field illumination. The growth of the individual colonies was quantitated using a computer-assisted photodensitometry method of determining the colonies' mass. The end buds showed the greatest growth response (greater than 20-fold increase in 3 weeks with a minimum doubling time of 48 h) to medium supplemented with 50% equine serum and cholera toxin or with less serum (15%) and epidermal growth factor (EGF), hormones and cholera toxin. Initially, the colonies grew logarithmically, then slowed as the colony size increased. This was due, in part, to the cessation of growth by the cells in the center. This end bud culture system differs from that of dissociated mouse mammary cells in a number of ways which are discussed.