流式细胞术测定淋巴细胞的分裂

Determination of Cell Division of Lymphocytes by Flow Cytometry

为确立流式细胞仪检测淋巴细胞的分裂的新方法 ,利用CFSE标记新鲜分离外周及胸腺淋巴细胞后 ,加入PMA、ConA、抗TCR抗体等培养 3d ,染色进行FACS分析。结果 ,诱导T细胞活化分裂的 3种刺激剂 ,以PMA刺激作用最强 ,ConA次之 ,抗TCR抗体最弱。表明CFSE标记细胞 ,流式细胞仪分析法不仅可以检测单细胞水平上细胞的分裂 ,还可根据荧光强度判断细胞的分裂次数。

To establish a new method to analyze the cell division capacity of murine T lymphocytes and thymocytes,fresh peripheral T lymphocytes and thymocytes were separated and stained with CFSE(5 carboxyfluorescein diacetate succinimidyl ester) These cells were then cultivated under stimulation by anti TCRαβ mAb, Con A and PMA ionomycin for 3 days, and subjected to analyze through FACS It was found that the effects of different stimuli on the cell division of T lymphocytes were different with the strongest stimulatory effect of PMA ionomycin; next of ConA and least of anti TCR mAb These results show that FACS analysis with CFSE staining permits both for the detection of cell division at single cell level and for the determination of numbers of cell division based on the strength of fluorescene