Establishment of Thymic Epitherial Cell Lines from a Human Thymoma and Its Immuno-biological Analysis.

人胸腺瘤胸腺上皮细胞系的建立及其免疫生物学分析。

基本信息

批准号：
06454392
负责人：
KOBAYASHI Shunsuke
金额：
$ 3.33万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454392/
关键词：
Establishment Cell line Thymic epitherial cell Thymoma in vitro in vivo Immunology Biology CD8 ヒト胸腺腫組織培養 cell line 免疫機能細胞生物学株化樹立 Subline

项目摘要

Using isolation culture techniques, we have been able to establish human thymic epitherial cells (Thm-1) from a thymoma of a 56 year-old female patient and have developed several subtype cell lines using cloning culture method. The growth characteristics of the cloning cell lines can be roughly divided into twotypes, polygonal and spindle types. Doubling time in vitro was approximately 1.3 day in polygonal Subline (3D) and 7.4 day in spindle Subline (7A). These cells were demonstrated to be highly tumorigenic when injected into nude mice. Thm-1 tumors, grown in nude mice, are histologically identical to the original tumor without lymphcytes. Immunophenotypic analysis of the cells using flow cytometry showed epitherial tumor cells expressing a surface E-cadherin and CEA antigen. Immunohistologically, the cells were demonstrated to express cytokeratin on the cell surface. Electron microscopy of the cultured cells revealed desmozome, tonofirament and granule particles. Nothern analysis demonstrated that E-cadherin mRNA was revealed in these cells. These data indicate that this cell line represents the epitherial lineage. To measure immunological reactivity, mixed lymphocyte-tumor cell culture reaction (MLTR) was studied using a microplate method. The PBL cells were cocultured with Thm-1 cells, lung cancer cell lines and macrophages in the medium supplemented with 200 mu/ml IL-2 and analyzed at periodic intervals for expanded lymph cell-subsets by flow cytometry.We found increased numbers of CD8 positive cells in 16-and 31-day cultures from the coculturing of Thm-1 cells and macrophages (58.7 after 16 days and 72.1% after 31 days of culture), but were small for CD4 positive cells (9.7% after 16 days and 7.5% after 31 days of culture).Establishment of this human thymoma cell line, Thm1, provides an excellent model to study further the biological behavior of thymic epitherial cells and the in vivo immunological effects of a thymoma.

使用隔离培养技术，我们能够从56岁女性患者的胸腺瘤中建立人类胸腺粘膜细胞（THM-1），并使用克隆培养方法开发了几种亚型细胞系。克隆细胞系的生长特性可以大致分为两种，多边形和主轴类型。在多边形subline（3D）和主轴subline（7a）中，体外时间约为1.3天，7.4天。当注射到裸鼠中时，这些细胞被证明是高度肿瘤的。在裸鼠中生长的THM-1肿瘤在组织学上与没有淋巴细胞的原始肿瘤相同。使用流式细胞术对细胞进行免疫表型分析显示表达表面E-钙粘着蛋白和CEA抗原的腹膜肿瘤细胞。在免疫组织学上，这些细胞被证明在细胞表面表达细胞角蛋白。培养细胞的电子显微镜揭示了脱菌病，牙皮病和颗粒颗粒。 Nothern分析表明，在这些细胞中揭示了E-钙粘蛋白mRNA。这些数据表明该细胞系代表层状谱系。为了测量免疫学反应性，使用微孔板方法研究了混合淋巴细胞肿瘤细胞培养反应（MLTR）。将PBL细胞与THM-1细胞，肺癌细胞系和巨噬细胞共培养，并在补充了200 mu/ml IL-2的培养基中，并以周期性的间隔进行分析，并通过流式细胞仪进行周期性的间隔，以通过流式细胞仪进行扩展的淋巴细胞群。我们发现，在16天和16天培养了16天的CD8阳性细胞的数量增加了16天和thm-1的16天（50天）（50天）（50天）。培养），但对于CD4阳性细胞的培养很小（16天后9.7％，培养31天后的7.5％）。建立这种人胸腺瘤细胞系THM1，提供了一个很好的模型，可以进一步研究胸腺瘤和胸腺瘤的体内免疫学作用的生物学行为。