U937巨噬细胞源性泡沫细胞模型的建立

Establishing a macrophage-derived foam cell model using U937cell lines

目的:建立稳定的人类单核细胞源性泡沫细胞模型,对研究动脉粥样硬化有重要的意义。实验拟以人类单核细胞株U937体外细胞培养的方法,模仿体内生理过程,建立一个新的人单核细胞源性泡沫细胞模型。方法:实验于2006-03/09在承德医学院基础医学研究所进行。①实验材料:实验中应用的U937细胞株由中科院细胞研究所提供。②实验方法:将16瓶密度为109L-1的U937细胞悬液分成对照组8瓶和实验组8瓶,实验组培养液中含100μg/L佛波酯、100mg/L低密度脂蛋白胆固醇,对照组细胞培养液中含100mg/L低密度脂蛋白胆固醇,不加佛波酯,置于37℃、体积分数为0.05的CO2、饱和湿度培养箱培养72h。③实验评估:收集细胞涂片采用油红O染色观察细胞形态,并测定细胞内总胆固醇含量。结果:实验组细胞内可见大量油红O染色阳性的脂质颗粒,对照组细胞形态无变化,细胞内无脂质颗粒。实验组细胞内总胆固醇高于对照组,两组差异具有显著性意义[(520.13±37.52),(39.47±9.26)mg/g,t=35.18,P<0.001]。结论:U937巨噬细胞源性泡沫细胞模型建立成功。

AIM： Stable human macrophage-derived foam cell model is significant for the study on artherosclerosis. This study investigated the feasibility of establishing macrophage-derived foam cell model using U937 cell lines. METHODS： The experiment was performed at Institute of Basic Medicine, Chengde Medical College from March to September 2006. （1）U937 cell lines were purchased from Institute of Biochemistry ＆ Cell Biology, Chinese Academy of Sciences. （2）Sixteen bottles of U937 cells （10^9 L^-1） were incubated at 37 ℃ in saturated humidity containing 5% CO2 for 72 hours. Among them, eight bottles contained 100 μg/L phorbol-12-myristate-13-acetate （PMA） and 100 mg/L low-density lipoprotein （LDL） as experimental group, and the other eight bottles only 100 mg/L LDL as control group. （3）Cell morphology was studied under light microscope by Wright＇s and Oil red O staining. Cell total cholesterol （TC） was measured after 72 hours of incubation. RESULTS： A large amount of lipid droplets were found in the cytoplasm by Oil red O staining in cells of the experimental group, but not found in control group cells. TC in cells of the experimental group was significantly higher than in control group [（520.13±37.52）, （39.47±9.26） mg/g, t=-35.18, P 〈 0.001]. CONCLUSION： Macrophage-derived foam cell model is established successfully in vitro using U937cells.