CHARACTERIZATION INVITRO OF LUMINAL AND MYOEPITHELIAL CELLS ISOLATED FROM THE HUMAN MAMMARY-GLAND BY CELL SORTING
CHARACTERIZATION INVITRO OF LUMINAL AND MYOEPITHELIAL CELLS ISOLATED FROM THE HUMAN MAMMARY-GLAND BY CELL SORTING
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DOI：
10.1111/j.1432-0436.1991.tb00883.x
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发表时间：
1991-04-01
期刊：
影响因子：
2.9
通讯作者：
GUSTERSON, BA
中科院分区：
文献类型：
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作者：
OHARE, MJ;ORMEROD, MG;GUSTERSON, BA
已使用荧光激活细胞分选从正常成人乳腺上皮中分离出管腔和肌上皮细胞。 它们的分离是基于两种表面抗原，上皮膜抗原（EMA）和常见的急性淋巴细胞白血病抗原（CALLA/CD 10/中性内肽酶24.11）的排他性表达。 分选的管腔和肌上皮细胞显示出明显不同的形态时，保持在单层培养，差异增强通过添加氢化可的松，胰岛素和霍乱毒素的培养基。 EMA阳性细胞形成了一个衰减的单层，细胞边界不清楚，而CALLA阳性细胞，相比之下，形成了紧密排列的可触知细胞阵列。 细胞类型特异性标记物细胞角蛋白18（管腔细胞）和平滑肌α-肌动蛋白（肌上皮细胞）的分布表明分选的群体约为98%纯。 然而，一个显着的少数（约15%）的分选管腔细胞一致表达的基底细胞标记细胞角蛋白14的文化。 一个显着的差异，注意到在增殖行为的两种类型的分选细胞，与肌上皮细胞迅速分裂，响应于体液添加剂，在相反的管腔细胞增殖缓慢。 两种类型的分选细胞可以在小鼠成纤维细胞饲养层的存在下克隆。 管腔和肌上皮细胞的克隆也是独特的;所有“传播”管腔克隆在外观上彼此相似，但在“紧凑”肌上皮克隆中观察到一些细胞异质性，包括鳞状化生。 这两种类型被证明保留了其原始的表面标记，并表现出不同的细胞骨架抗原表型时，他们重新分析后3周的生长期。 当分别分离导管和肺泡进行克隆时，获得了扩展和紧凑的表型。 通过流式细胞术从人乳腺上皮中分离的细胞的详细表征为进一步研究体外纯化细胞类型的管腔-肌上皮相互作用和生长反应提供了基础。
Luminal and myoepithelial cells have been separated from normal adult human breast epithelium using fluorescence activated cell sorting. Their isolation was based on the exclusive expression of two surface antigens, epithelial membrane antigen (EMA) and the common acute lymphoblastic leukaemia antigen (CALLA/CD10/neutral endopeptidase 24.11). Sorted luminal and myoepithelial cells displayed distinctively different morphologies when maintained in monolayer culture, differences which were enhanced by the addition of hydrocortisone, insulin and cholera toxin to the culture medium. The EMA-positive cells formed an attenuated monolayer with indistinct cell boundaries while CALLA-positive cells, by contrast, formed tightly packed arrays of refractile cells. The distribution of the cell type-specific markers cytokeratin 18 (luminal cells) and smooth muscle alpha-actin (myoepithelial cells) indicated that the sorted populations were approximately 98% pure. However, a significant minority (approximately 15%) of sorted luminal cells consistently expressed the basal-cell marker cytokeratin 14 in culture. A marked difference was noted in the proliferative behaviour of the two types of sorted cells, with myoepithelial cells dividing rapidly in response to the humoural additives, in contrast to the luminal cells which proliferated slowly. Both types of sorted cells could be cloned in the presence of feeder layers of mouse fibroblasts. Clones of luminal and myoepithelial cells were also distinctive; all "spread" luminal clones were similar in appearance to each other, although some cellular heterogeneity, including squamous metaplasia, was observed in "compact" myoepithelial clones. Both types were shown to have retained their original surface markers and to exhibit different cytoskeletal antigenic phenotypes when they were re-analysed after a 3-week growth period. Both spread and compact phenotypes were obtained when separately isolated ducts and alveoli were cloned. This detailed characterization of cells isolated from the human breast epithelium by flow cytometry provides the basis for further studies of luminal-myoepithelial interactions and growth responses of purified cell types in vitro.