| Project/Area Number |
06454392
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | Tohoku University |
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Principal Investigator |
KOBAYASHI Shunsuke Tohoku University, Institute of Deveropment, Aging and Cancer, Associate Professor, 加齢医学研究所, 助教授 (00125543)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1994: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | 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 |
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| Research Abstract |
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.
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