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G CSF体外诱导新生小鼠心肌干细胞分化的研究 |
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【摘要】 目的 了解新生小鼠心肌干细胞(cardiac stem cells,CSCs)在体外的分化潜能,为重建缺血及坏死心肌的临床应用提供依据。方法 采用组织块贴壁法体外培养小鼠CSCs,以免疫磁珠细胞分离 (MACS)系统纯化原代细胞。纯化所得细胞分别予4个浓度的粒细胞集落刺激因子(GCSF)进行诱导,应用流式细胞术(FCM)、免疫荧光染色(IFS)鉴定细胞表型,RTPCR检测心肌转录因子GATA4与心肌结构基因βMHC的表达。结果 由组织块培养所得的原代细胞经FCM、IFS检测证实其部分表达Sca1 和CD 45。采用MACS成功得到高纯度的细胞,行FCM、IFS鉴定其表型为Sca1+/CD 45-,RTPCR检测显示其表达GATA4,不表达βMHC。纯化的细胞予诱导后培养3周,整个培养过程未见单个或成团细胞的自发搏动。诱导后3周各组细胞再次经FCM检测见Sca1的表达均比诱导前减弱,RTPCR检测见诱导后的细胞仍然不表达βMHC。结论 应用MACS后成功得到高纯度的表型为Sca1+/CD 45-的CSCs;CSCs Sca1的表达量可能随培养时间的延长而下降;GCSF未能将CSCs诱导分化为心肌细胞。
【关键词】 心肌干细胞; 粒细胞集落刺激因子; 心肌再生
Differentiation of Cardiac Stem Cells Induced by GCSF in Vitro
YANG Zhenjun,LU Dongfeng,OU Nianwen,HU Dexi
(Department of Cardiology,Second Affiliated Hospital of Guangzhou Medical College,Guagnzhou 510260,China.)
Abstract: Objective To investigate the differentiate potential of cardiac stem cells (CSCs)isolated from neonatal mice,and provide a scientific basis for the clinical application to reconstruct ischemic and necrotic myocardium.Methods CSCs were cultured in vitro and were purified by magnetic cell separation (MACS)system.The purified cells were respectively induced with four different concentrations of granulocyte colonystimulating factor (GCSF)Flow cytometry (FCM)and immunofluorescence staining (IFS)were performed to identify cell phenotype.Reve[1] [2] [3] [4] [5] [6] [7] [8] [9] 下一页 上一个医学论文: 别嘌呤醇治疗伴高尿酸血症的慢性心力衰竭疗效观察 下一个医学论文: 体外血小板流动腔研究方法的建立和应用
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