ESTABLISHMENT AND CHARACTERIZATION OF MURINE MACROPHAGE-LIKE CELL-LINES FOLLOWING TRANSFORMATION WITH SIMIAN-VIRUS-40 DNA DELETED AT THE ORIGIN OF REPLICATION
ESTABLISHMENT AND CHARACTERIZATION OF MURINE MACROPHAGE-LIKE CELL-LINES FOLLOWING TRANSFORMATION WITH SIMIAN-VIRUS-40 DNA DELETED AT THE ORIGIN OF REPLICATION
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DOI：
10.1016/0022-1759(94)90008-6
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发表时间：
1994-09-14
影响因子：
2.2
通讯作者：
MACDONALD, C
中科院分区：
文献类型：
--
作者：
KREUZBURGDUFFY, UCK;MACDONALD, C
通过将病毒致癌基因导入原代细胞，可以建立分化的哺乳动物细胞系。这类细胞系在适应长期文化生活的同时，还能保留其原有的专门功能；但是，用这种方法获得的大多数转化细胞系与原代细胞相比，其特性都发生了改变。这些变化的结果是，转化的细胞系不再提供原始组织的良好模型，实际上往往比初始细胞类型更像其他转化的细胞系。在我们的实验室中，通过将原代细胞转化为SV40来源缺失的DNA，分离了三种小鼠腹膜巨噬细胞样细胞系。这些细胞系在连续培养中大约1年，并在此期间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。细胞吞噬igg包被颗粒，小鼠巨噬细胞特异性标志物F4/80阳性，具有抗原呈递功能。溶菌酶、酸性磷酸酶、纤溶酶原激活剂、胶原酶、前列腺素E2和5′-核苷酸酶活性也在这些品系中检测到。在本文中，DNA转化的方法将被描述，以及一些用于表征三个永生化细胞系的测定。
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.