Isolation of the epithelial subcomponents of the mouse mammary gland for tissue-level culture studies
Isolation of the epithelial subcomponents of the mouse mammary gland for tissue-level culture studies
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分离小鼠乳腺上皮亚成分用于组织水平培养研究
DOI：
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发表时间：
1983
期刊：
影响因子：
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通讯作者：
R. Dils
中科院分区：
文献类型：
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作者：
W. Jones;R. Hallowes;N. Choongkittaworn;H. Hosick;R. Dils
我们开发了一种用于小鼠乳腺的组织解离程序，由此可以将实质分离为基本上不含间充质的完整结构子成分。将整个乳腺脂肪垫粗略切碎，并进行有限的胶原酶消化，在 37°C 下机械解离 90 分钟。实质以多细胞类器官和单分散细胞的混合物形式释放。使用分级系列过滤器（400、250、150、95 和 51 μ 孔径）可以将实质与非实质材料分离，并进一步将前者富集为导管、导管小叶和末端芽或肺泡群体。产量是可变的并且取决于起始组织的性质，例如小鼠品系、年龄和胎次。这些类器官级分可以在培养物中单独建立。该程序允许在组织水平上研究小鼠乳腺的导管和末端区域。
We have developed a tissue dissociation procedure for the mouse mammary gland whereby it is possible to isolate the parenchyma as intact structural subcomponents essentially free of mesenchyme. Whole mammary fat pads are coarsely minced and subjected to a limited collagenase digestion, with mechanical dissociation for 90 min at 37° C. The parenchyma is released as a mixture of multicellular organoids and monodispersed cells. Use of a graded series of filters (400, 250, 150, 95, and 51 μ pore size) allows the separation of parenchyma from nonparenchymal material, with a further enrichment of the former into ductal, ductal-lobular, and terminal end-bud or alveolar populations. Yield is variable and dependent upon the nature of the starting tissue, e.g., mouse strain, age, and parity. These organoid fractions may be established separately in culture. This procedure allows for the study of the ductal and terminal regions of the mouse mammary gland in culture at the tissue level.