EPO工程细胞株支原体检测

THE EXAMINATION FOR MYCOPLASMA IN EPO C_2 ENGINEERING CELL LINE

支原体污染是细胞培养过程中最常见的问题之一,对细胞支原体的检测是细胞特性鉴定的重要方面。我们采用抗支原体单抗免疫荧光法和培养法(包括液体培养法和固体培养法)检测工程细胞支原体。单抗免疫荧光法检测结果表明:支原体阳性的细胞膜表面有明亮荧光,工程细胞EPO C_2细胞膜表面无荧光。支原体液体培养法结果显示:作为阳性对照的支原体由于在生长过程中产酸使培养液变黄,而加入EPOC_2株细胞悬液的样品管与阴性对照管相同,无颜色变化。固体培养法结果显示:在显微镜下观察,支原体阳性对照在固体培养基上呈荷包蛋样集落,而阴性对照及EPO C_2株细胞样品均无菌落生长。以上结果表明:受检的EPO C_2株细胞未被支原体污染。

Mycoplasma contamination raust be prevented during cell culture. Mycoplasma examination is an importam as-pect in determinating the characteristics of engineering cell line. There are many methods for detecting mycoplas-rna .including PCR method,probe blot method,culture methods(liquid and solid culture)and so on. In this study,we examine mycoplasma in EPO C2 engineering cell line with an immunofluorescence method in which we use mono-clonal antibody as probe. The results show that the EPO C2 cell line examined has not been contaminated by my-coplasma.