Péchoux C, Gudjonsson T, Ronnov-Jessen L, Bissell M J, Petersen O W
Department of Anatomy, Section A, The Panum Institute, University of Copenhagen, Blegdamsvej 3, Copenhagen N, DK-2200, Denmark.
Dev Biol. 1999 Feb 1;206(1):88-99. doi: 10.1006/dbio.1998.9133.
The origin of the epithelial and myoepithelial cells in the human breast has not been delineated. In this study we have addressed whether luminal epithelial cells and myoepithelial cells are vertically connected, i.e., whether one is the precursor for the other. We used a primary culture assay allowing preservation of basic phenotypic traits of luminal epithelial and myoepithelial cells in culture. The two cell types were then separated immunomagnetically using antibodies directed against lineage-specific cell surface antigens into at best 100% purity. The cellular identity was ascertained by cytochemistry, immunoblotting, and 2-D gel electrophoresis. Luminal epithelial cells were identified by strong expression of cytokeratins 18 and 19 while myoepithelial cells were recognized by expression of vimentin and alpha-smooth muscle actin. We used a previously devised culture medium (CDM4) that allows vigorous expansion of proliferative myoepithelial cells and also devised a medium (CDM6) that allowed sufficient expansion of differentiated luminal epithelial cells based on addition of hepatocyte growth factor/scatter factor. The two different culture media supported each lineage for at least five passages without signs of interconversion. We used parallel cultures where we switched culture media, thus testing the ability of each lineage to convert to the other. Whereas the myoepithelial lineage showed no signs of interconversion, a subset of luminal epithelial cells, gradually, but distinctly, converted to myoepithelial cells. We propose that in the mature human breast, it is the luminal epithelial cell compartment that gives rise to myoepithelial cells rather than the other way around.
人类乳腺上皮细胞和肌上皮细胞的起源尚未明确。在本研究中，我们探讨了管腔上皮细胞和肌上皮细胞是否垂直相连，即一种细胞是否是另一种细胞的前体。我们采用了一种原代培养试验，该试验能够在培养过程中保留管腔上皮细胞和肌上皮细胞的基本表型特征。然后，使用针对谱系特异性细胞表面抗原的抗体，通过免疫磁珠法将这两种细胞类型分离，纯度最高可达100%。通过细胞化学、免疫印迹和二维凝胶电泳确定细胞身份。通过细胞角蛋白18和19的强表达鉴定管腔上皮细胞，而波形蛋白和α-平滑肌肌动蛋白的表达则可识别肌上皮细胞。我们使用了一种先前设计的培养基（CDM4），该培养基能够使增殖性肌上皮细胞大量扩增，还设计了一种基于添加肝细胞生长因子/分散因子的培养基（CDM6），该培养基能够使分化的管腔上皮细胞充分扩增。这两种不同的培养基支持每个谱系至少传代五次，且没有相互转化的迹象。我们进行了平行培养，并在培养过程中更换培养基，从而测试每个谱系转化为另一个谱系的能力。虽然肌上皮谱系没有相互转化的迹象，但一部分管腔上皮细胞逐渐但明显地转化为肌上皮细胞。我们提出，在成熟的人类乳腺中，是管腔上皮细胞产生了肌上皮细胞，而不是相反。
