EB病毒相关淋巴母细胞样细胞系的建立及鉴定

Development and identification of lymphoblastoid cell lines associated with Epstein-Barr virus

目的由于EB病毒与肿瘤的关系极为复杂且存在争议,淋巴母细胞样细胞系LCLs(lymphoblastoid celllines)没有肿瘤形成过程中的复杂因素,从而成为了研究EBV介导的细胞永生化的最佳模型。本实验旨在建立EBV原型株相关的永生化淋巴母细胞系,为后期LCLs的端粒酶活性及其体内外成瘤性的相关研究提供基础。方法利用体外培养B95.8细胞产生EB病毒原型株感染正常人外周血淋巴细胞培养传代得到5株EB病毒相关淋巴母细胞,并用RT-PCR对其进行了基因表达、流式细胞仪进行细胞表面标记以及核型的鉴定。结果淋巴母细胞样细胞系能不断传代,能表达EBV基因,具有成熟B细胞表面标记;细胞传代过程早期未发生核型改变。结论建立了5株永生化细胞株,分别为LCL1-5、LCL2-5、LCL3-5、LCL4-30、LCL5-30。

Objective The relation between Epstein-Barr virus and tumor is complicated and disputable. Lymphoblastoid cell lines （LCLs） have become the best model in the studies on tumorgenesis and cell immortalization associated with EBV because of its relatively simple process, furthermore some characters of B lymphocyte make LCLs important in the researches in immunology. In this paper, we aimed to develop LCLs associated with EBV to carry on our following experiments on the telomerase activity and tumorgenesis in vivo and in vitro. Methods Normal peripheral blood lymphocytes were transformed with EBV prototype produced by B95.8 cell; LCLs were identified in gene expression of EBV by RT-PCR, and cell surface marker and karyotype were also detected. Results 5 strains of LCLs could be subcultured continently with normal karyote, EBV associated latent membrane protein gene and CD19 could be dected in all of the five cell lines. Conclusion five strains of LCLs were developed and named respectively with LCL1-5, LCL2-5, LCL3-5, LCL4-30,LCL5-30.