Clarke C, Titley J, Davies S, O'Hare M J
Institute of Cancer Research, Haddow Laboratories, Sutton, Surrey, UK.
Epithelial Cell Biol. 1994 Jan;3(1):38-46.
A comparison has been made between different immunomagnetic techniques for separating normal human mammary epithelial cells based on the exclusive expression of EMA (epithelial membrane antigen) by luminal cells and CALLA (CD10) by myoepithelial cells. When cells labelled with antibodies to these antigens were incubated with Dynabeads, they rosetted myoepithelial but not luminal cells. However, both luminal and myoepithelial cells could be positively separated using the MACS system. Purity was established by analyzing the expression of CALLA and EMA using flow cytometry, and of cell-type specific cytokeratins using indirect immunofluorescence. Dynabead-separated myoepithelial cell populations were of high purity (> 98%) but the beads could not be removed from the cells. Luminal cell populations separated by the MACS method were also highly purified (> 95%), as were myoepithelial cell populations (> 90%). Using this immunomagnetic separation method, up to 10(7) cells of each type could be obtained from individual preparations.
基于管腔细胞特异性表达上皮膜抗原（EMA）以及肌上皮细胞特异性表达CALLA（CD10），对不同免疫磁珠技术分离正常人乳腺上皮细胞的效果进行了比较。当用针对这些抗原的抗体标记细胞，并与磁珠孵育时，磁珠会与肌上皮细胞形成玫瑰花结，但不会与管腔细胞形成玫瑰花结。然而，使用MACS系统可以将管腔细胞和肌上皮细胞都进行阳性分离。通过流式细胞术分析CALLA和EMA的表达，以及使用间接免疫荧光分析细胞类型特异性细胞角蛋白的表达来确定纯度。磁珠分离的肌上皮细胞群体纯度很高（>98%），但磁珠无法从细胞中去除。通过MACS方法分离的管腔细胞群体也高度纯化（>95%），肌上皮细胞群体也是如此（>90%）。使用这种免疫磁珠分离方法，每次制备可从个体样本中获得多达10⁷个每种类型的细胞。
