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

发布时间：2018-07-14 12:45

【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。
[Abstract]:Objective to study the method of isolation and culture of mouse thymus epithelial cells and its proliferative effect on thymocytes. Methods the thymus of BALB / c mice aged 4 weeks and 5 weeks old were collected and cultured for 30 days. The changes of cell morphology in different time were observed and the digestive cells were subcultured with 0.1% trypsin, and identified by immunohistochemistry with vimentin and keratin. The inhibitory effects of dexamethasone on thymocytes, spleen lymphocytes and coculture of thymus lymphocytes and thymoepithelial cells were detected by colorimetric assay. Results on the 3rd day of culture, a few thymus epithelial cells were found around the adherent tissue mass. The cells were round, polygonal, dendritic, fusiform and so on from the inside to the outside, and the cells grew faster when cultured for about 10 days. A large number of adherent cells appeared; after 21 days of culture, the cells fused in large numbers and presented a typical cobblestone arrangement. Cell immunohistochemistry showed that broad-spectrum keratin was positive, vimentin was negative, and the thymus gland was increased with the increase of drug concentration. The survival rate of spleen lymphocytes decreased and the survival rate of thymus lymphocytes increased after co-culture. Conclusion it is simple and feasible to culture mouse thymoepithelial cells by tissue mass method, and thymoepithelial cells can promote the proliferation of thymocytes.
【作者单位】：广东医学院生物学教研室;
【基金】：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)
【分类号】：R329.2

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