| 小鼠胸腺上皮细胞的培养、鉴定及对淋巴细胞促增殖作用的初步研究 |
| 投稿时间：2014-09-10 修订日期：2014-09-26 点此下载全文 |
| 引用本文：郑丹丹,欧阳军,何滔,张华华.小鼠胸腺上皮细胞的培养、鉴定及对淋巴细胞促增殖作用的初步研究[J].医学研究杂志,2015,44(3):41-44 |
| DOI：
10.3969/j.issn.1673-548X.2015.03.012 |
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| 基金项目:国家自然科学基金资助项目(31200679);广东省科技计划项目(2011B031800388);广东省大学生创新训练项目(1057112027) |
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| 中文摘要:目的 研究小鼠胸腺上皮细胞的分离培养方法、鉴定及其对胸腺淋巴细胞的促增殖作用。方法 无菌取4~5周龄BALB/c小鼠的胸腺, 组织块培养30天, 观察不同时间细胞形态的变化;0.1%胰蛋白酶消化细胞传代;用波形蛋白和角蛋白进行免疫组化鉴定;新型细胞增殖及细胞毒性检测试剂盒比色法检测地塞米松对胸腺、脾脏淋巴细胞单独培养及胸腺淋巴细胞与胸腺上皮细胞共培养的抑制作用。结果 培养至第3~4天可见贴壁的组织块周围出现少量胸腺上皮细胞, 从内到外细胞形状为圆形、多边形、树突状、梭形等;培养至10天左右时, 细胞生长较快, 出现大量的贴壁细胞;培养21天左右细胞大量融合并呈典型的鹅卵石样排列;细胞免疫组化显示广谱角蛋白呈阳性反应, 波形蛋白呈阴性反应;随着药物浓度的增加, 胸腺、脾脏淋巴细胞存活率均降低, 共培养后胸腺淋巴细胞的存活率升高。结论 组织块法体外培养小鼠胸腺上皮细胞简单可行;胸腺上皮细胞具有促胸腺淋巴细胞增殖的作用。 |
| 中文关键词:小鼠 胸腺上皮细胞 胸腺淋巴细胞 |
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| Culture and Identification of Mouse Thymic Epithelial Cells and Preliminary Study on Promoting Lymphocyte Proliferation |
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| Abstract:Objective To study the method of isolation and culture of mouse thymic epithelial cells (TEC), the identification of its role in the thymic lymphocyte proliferation. Methods The thymus from 4-5 weeks old BALB/c mice were obtai ned under sterile conditions. The change of the cell morphology at different time was cultured for 30 days and observed. Thymic epithelial cells were passaged using 0.1% trypsin and immunohistochemistry identified by anti-vimentin and anti-keratin. The effect of dexamethasone on proliferation activity of thymus and spleen lymphocyte was detected.They were cultured alone and co-cultured with thymocytes and thymic epithelial cells with cell counting kit-8 colorimetric assay. Results A small number of thymic epithelial cells appeared surrounding adherent tissue block at 3-4 days, and shaped circular, polygon, spindle, etc, and the residual lymphocyte can be removed by the replacement of medium gradually.About 10 days cells grow faster and a large numberof adherent cells appeared. A large number of cell fused and showed a typical cobblestone pattern about 21 days. Passaged cells were polygonal and cell morphology was not clearer with the primary cells and immunohistochemistry showed anti-keratin positive and anti-vimentin negative. With the increasing of drug concentration, thymus, spleen lymphocyte survival rates were lower, and the survival rate of thymus lymphocyte increased after co-cultured with thymic epithelial cells. Conclusion Tissue cultured method of mousethymic epithelial cells in vitro is simple and feasible and thymic epithelial cells can promote the thymus lymphocytes proliferation. |
| keywords:Mouse Thymic epithelial cells Thymic lymphocytes |
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