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DMSO诱导HL-60转化为类中性粒细胞的实验研究
Research of Experiment on Neutrophil-like Cells Differentiated from HL-60 Induced by DMSO
【摘要】 通过DMSO诱导HL-60细胞分化为类中性粒细胞,从细胞形态、CD11b基因的表达等方面进行研究,探索这种类中性粒细胞是否可以作为进一步研究中性粒细胞自发性凋亡死亡机制的细胞模型。体外培养HL-60细胞,在对数生长期细胞加入DMSO(终浓度1%)对其进行刺激和诱导,然后分别于24、48、72、144 h收集细胞,以未加DMSO处理的相应各时间点HL-60细胞作为空白对照组,运用光学显微镜镜下观察细胞的形态学变化,用RT-PCR检测分化标志物CD11b基因的表达。结果显示:随着诱导时间的延长被诱导细胞的体积逐渐增大,48 h后出现了肾形核、分叶核、蚕豆核等粒细胞核型,并随时间的延长逐渐增多;DMSO处理后,24 h开始类中性粒细胞中CD11b的表达上调,与空白对照组相比有明显差异(P<0.01)。实验结果表明:DMSO可以诱导HL-60细胞分化为类中性粒细胞;类中性粒细胞具有成为中性粒细胞研究替代模型的潜能。
【Abstract】 Here we investigated HL-60 cell induced by dimethylsulphoxide(DMSO) to differentiate into neutrophil-like cells.HL-60 cell was cultivated in vitro and induced by DMSO(final concentration 1%) at exponential phase.HL-60 cells stimulated by DMSO were used as experimental group.HL-60 cells without stimulation were used as negative control.Cell morphology of different cultured conditions was observed by optical microscope.CD11b mRNA expression of different groups was detected by RT-PCR.Results showed that the bulk of induced cells are larger and larger as induced time goes by.The caryomorphism looks like lobulated,reniform and fabaceous.The result of RT-PCR showed that the expression of CD11b mRNA was up-regulated comparing with the negative control groups since DMSO 24 hours(P<0.01).All data suggest that neutrophil-like cells can be derived from HL-60 cells induced by DMSO,and neutrophil-like cells could be the substitution model for neutrophil research.
- 【文献出处】 重庆理工大学学报(自然科学) ,Journal of Chongqing University of Technology(Natural Science) , 编辑部邮箱 ,2011年08期
- 【分类号】R392.11
- 【被引频次】3
- 【下载频次】774