Analysis of gene expression in FACS-sorted thymocytes

FACS 分选胸腺细胞的基因表达分析

• 批准号: 13670002
• 负责人: ITOH Tsunetoshi
• 金额: $ 2.69万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670002/
• 关键词: Thymus; Thymocytes; Cell sorter; Selection; Differentiation; Surface marker; Gene expression; TCR; セルソーティング; 同定

Thymocytes, derived from hematopoietic stem cells in the bone marrow, proliferate and differentiate into mature T cells after entering the thymus. One of the biggest problems for thymocyte differentiation is that the accurate selection mechanism is still unresolved. How T cell receptor is closely and chronologically interrelated with "the selection process", how self-reactive clones are eliminated, and how CD4 and CD8 are differentially expressed, are still uncertain.To address these issues, we believe that the important key should be the isolation and identification of the subset about to go through the selection process. In this project, experiments were designed to analyse gene expression in subsets just before or after the selection. On the other hand, since, we realized surface markers and the computer application capable of analyzing their expression are not sufficient and not well coordinated, we undertook the substantial improvement of the analyzing program. With this renewed application, we could identify several subsets not noticed so far but may have apparent significance in thymocyte differentiation.A fundamentally new pathway for thymocyte differentiation is now being progressively constructed, and the findings would be an important infomation to delineate and identify thymocyte subsets about to enter the critical selection process.

胸腺细胞来源于骨髓中的造血干细胞，进入胸腺后增殖并分化为成熟的T细胞。胸腺细胞分化的最大问题之一是准确的选择机制尚未解决。 T细胞受体如何与“选择过程”密切相关、时间顺序如何、自身反应性克隆如何被消除、CD4和CD8如何差异表达等，目前仍不确定。要解决这些问题，我们认为重要的关键应该是即将经历选择过程的亚群的分离和鉴定。在该项目中，实验旨在分析选择之前或之后子集中的基因表达。另一方面，由于我们意识到表面标记和能够分析其表达的计算机应用程序还不够充分且协调得不好，因此我们对分析程序进行了实质性改进。通过这一新的应用，我们可以识别出一些迄今为止尚未注意到但可能在胸腺细胞分化中具有明显意义的亚群。目前正在逐步构建胸腺细胞分化的全新途径，这些发现将成为描述和识别即将进入关键选择过程的胸腺细胞亚群的重要信息。

项目成果

Itoh, T., et al.: "Establishment of a mouse thymic epithelial cell line, IT-79MHC and a brief review on cultured thymic epithelial cells"Cellular and Molecular Biology. 47. 1-18 (2001)

Itoh, T. 等人：“小鼠胸腺上皮细胞系 IT-79MHC 的建立以及培养胸腺上皮细胞的简要回顾”细胞和分子生物学。

Itoh, T., et al.: "Establishment of a mouse thymic epithelial cell line, IT-76MHC and a brief review on cultured thymic epithelial cells"Cellular and Molecular Biology. 47. 1-18 (2001)

Itoh, T. 等人：“小鼠胸腺上皮细胞系 IT-76MHC 的建立以及培养胸腺上皮细胞的简要回顾”细胞和分子生物学。

Toyosaki-Maeda,T., Takano,H., Tomita,T., Tsuruta,Y., Maeda-Tanimura,M., Shimaoka,Y., Takahashi,T., IToh,T., Suzuki,R. and Ochi,T.: "Differentiation of monocytes into multinucleated giant bone-resorbing cells : two-step differentatiation induced by nurse-l

丰崎前田，T.，高野，H.，富田，T.，鹤田，Y.，前田谷村，M.，岛冈，Y.，高桥，T.，IToh，T.，铃木，R。

TamuraA., et al.: "Distribution of two types of lymphocytes, intraepithelial and lamina propria-associated ones, in the murine small intestine"Cell and Tissue Research. 311(In press). (2003)

TamuraA. 等人：“小鼠小肠中两种类型的淋巴细胞（上皮内淋巴细胞和固有层相关淋巴细胞）的分布”细胞和组织研究。

Tamura A. et al.: "Distribution of two types of lymphocytes, intraepithelial and lamina propria-associated ones, in the murine small intesine"Cell and Tissue Research. 311(in press). (2003)

Tamura A. 等人：“小鼠小肠中两种类型的淋巴细胞（上皮内淋巴细胞和固有层相关淋巴细胞）的分布”细胞和组织研究。