Kreuzburg-Duffy U C, MacDonald C
Department of Biological Sciences, University of Paisley, Scotland, UK.
J Immunol Methods. 1994 Sep 14;174(1-2):33-51. doi: 10.1016/0022-1759(94)90008-6.
Differentiated mammalian cell lines can be established by introducing viral oncogenes into primary cells. Such lines can retain their original specialised functions while being adapted to prolonged life in culture; but most transformed cell lines obtained in this way characteristically show altered properties compared with the primary cells. The result of these changes is that transformed cell lines no longer provide a good model of the original tissue, and indeed often resemble other transformed lines more than the initial cell type. In our laboratory three murine peritoneal macrophage-like cell lines have been isolated by transforming primary cells with SV40 origin-deleted DNA. These lines have been in continuous culture for approximately 1 year and have been shown to express many macrophage-specific properties throughout this time, including Fc receptors and staining for non-specific esterase. The cell lines phagocytosed IgG-coated particles, they were positive for the murine macrophage-specific marker F4/80 and they showed antigen-presentation function. Lysozyme, acid phosphatase, plasminogen activator, collagenase, prostaglandin E2 and 5'-nucleotidase activities have also been detected in these lines. In this paper the method of DNA transformation will be described as well as some of the assays used for the characterization of the three immortalized cell lines.
通过将病毒癌基因导入原代细胞，可以建立分化的哺乳动物细胞系。这样的细胞系在适应培养中的长期存活时，能够保留其原有的特殊功能；但通过这种方式获得的大多数转化细胞系与原代细胞相比，其特性会发生改变。这些变化的结果是，转化细胞系不再能很好地代表原始组织，实际上它们通常与其他转化细胞系的相似程度超过了初始细胞类型。在我们实验室中，通过用缺失SV40原点的DNA转化原代细胞，分离出了三种小鼠腹膜巨噬细胞样细胞系。这些细胞系已经连续培养了大约1年，并且在此期间一直表现出许多巨噬细胞特异性特性，包括Fc受体和非特异性酯酶染色。这些细胞系吞噬IgG包被的颗粒，对小鼠巨噬细胞特异性标志物F4/80呈阳性，并且具有抗原呈递功能。在这些细胞系中还检测到了溶菌酶、酸性磷酸酶、纤溶酶原激活物、胶原酶、前列腺素E2和5'-核苷酸酶的活性。本文将描述DNA转化方法以及用于表征这三种永生化细胞系的一些检测方法。
