Dundas S R, Ormerod M G, Gusterson B A, O'Hare M J
Institute of Cancer Research, Royal Cancer Hospital, Haddow Laboratories, Sutton, Surrey, UK.
J Cell Sci. 1991 Nov;100 ( Pt 3):459-71. doi: 10.1242/jcs.100.3.459.
Differentially expressed membrane antigens have been used to flow-sort viable luminal epithelial and myoepithelial cells from freshly disaggregated adult virgin rat mammary parenchyma. Resulting cultures and clones have been characterized morphologically and by a panel of antibodies that recognise cell-type-specific cytoskeletal antigens in the intact mammary gland. Five clonal phenotypes were recognisable by morphological criteria, three (types 1-3) exclusively associated with sorted luminal epithelial (25.5-positive) cells, and two (types 4 and 5) generated from the sorted myoepithelial (CALLA/neutral endopeptidase 24.11-positive) cells. All clones derived from myoepithelial cells continued to express a basal parenchymal marker in the form of the rat equivalent of human cytokeratin 14, while smooth muscle alpha-actin was expressed by the small slowly growing type 4 clones but was found in fewer cells in rapidly proliferating type 5 myoepithelially derived clones. Two of the luminal clone types, characterized by an attenuated appearance and slow growth (types 1/2), expressed only luminal-specific markers, including the equivalent of human cytokeratins 7/18/19. Type 3 clones, by contrast, consisted of rapidly proliferating cells, many of which either co-expressed CK14 and CK18 antigens (type 3a) or were composed of a mosaic of CK14+/CK18-, CK14+/CK18+ and CK14-/CK18+ cells (type 3b). The sorted myoepithelially derived clones grew faster than clones from sorted luminal cells as evidenced by the larger fraction of cells synthesizing DNA. All types of clone could be obtained from both isolated ducts and alveoli, when these were cloned separately, although there were some differences in their relative frequency.(ABSTRACT TRUNCATED AT 250 WORDS)
差异表达的膜抗原已被用于从刚解离的成年未孕大鼠乳腺实质中流式分选活的管腔上皮细胞和肌上皮细胞。对所得培养物和克隆进行了形态学表征,并通过一组抗体进行鉴定,这些抗体可识别完整乳腺中细胞类型特异性的细胞骨架抗原。通过形态学标准可识别出五种克隆表型,其中三种(1 - 3型)仅与分选的管腔上皮(25.5阳性)细胞相关,另外两种(4型和5型)由分选的肌上皮(CALLA/中性内肽酶24.11阳性)细胞产生。所有源自肌上皮细胞的克隆继续以大鼠等效的人细胞角蛋白14的形式表达基底实质标志物,而平滑肌α - 肌动蛋白由缓慢生长的小4型克隆表达,但在快速增殖的5型肌上皮来源克隆中的细胞中较少发现。两种管腔克隆类型,其特征为外观衰减和生长缓慢(1/2型),仅表达管腔特异性标志物,包括等效的人细胞角蛋白7/18/19。相比之下,3型克隆由快速增殖的细胞组成其中许多细胞共表达CK14和CK18抗原(3a型)或由CK14 + /CK18 - 、CK14 + /CK18 + 和CK14 - /CK18 + 细胞的镶嵌体组成(型)。分选的肌上皮来源克隆比分选的管腔细胞克隆生长得更快,这由合成DNA的细胞比例更大证明。当单独克隆时,所有类型的克隆都可以从分离的导管和肺泡中获得,尽管它们的相对频率存在一些差异。(摘要截断于250字)