CHO细胞微核的激光扫描细胞仪自动化检测方法

A primary study on automatic detection of micronucleus in CHO cells by laser scanning cytometry

运用激光扫描细胞仪,采用FITC和PI两种荧光染料,结合细胞胞质分裂阻断微核技术,对环磷酰胺(CP)诱导的中国仓鼠卵巢(CHO)细胞微核进行了检测,并将该方法与传统微核显微镜检法进行了比较,旨在探索CHO细胞体外微核的激光扫描细胞仪自动化检测方法.结果表明,激光扫描细胞仪方法能够准确识别胞质阻滞的双核细胞及双核细胞中的微核,环磷酰胺(CP)处理浓度与其诱导的双核细胞微核率成正相关,激光扫描细胞仪法与传统显微镜检法获得的微核率具有良好的相关性.激光扫描细胞仪检测方法能简便、经济、快速、高效地检测CHO细胞微核,适于体外培养细胞尤其是贴壁细胞微核的高通量检测.

To study a automatic detecton of micronucleus in Chinese hamster ovary （CH0） cells in vitro by laser scanning cytometry（ LSC）, the micronucleus of CHO cells induced by cyclophosphs.mide （CP） were detected by LSC eonbined with cytokinesis block micronucleus assay using two kinds of fluorescent dyes of fluorescein isothiocyanate （ FITC, a protein - specific dye） and propidium iodide （ PI, a DNA - specific dye）. Also, the micronucleus frequencies of CHO detected by LSC were compared with that calculated by a microscope here. Binucleated ceils and mieronucleus in the binucleated ceils can be identified by LSC accurately;and the micronucleus frequencies of binucleated cell induced by CP in the cytokinesis block micronucleus assay were found to be significantly higher than the control. Besides, the frequencies of micronucleus in CHO eells were increased along with higher concentration of cyclophosphamide. And the results of in vitro micronucleu, scored by LSC have a good correlation with that by the traditional microscopy. LSC is an easily operated method, and it detects micronucleus in CHO cells rapidly and accurately, as well as economically. It is suitable for detecting micronucleus of in vitro cells, especially for attachment cells.