心肌梗死大鼠体外培养心肌组织对间充质干细胞迁移的作用  

Effects of cardiac tissue mass cultured ex vivo on migration of mesenchymal stem cells in myocardial infarction rats

作　　者：王丁琼[1] 史若飞[2] 张晓刚[1] 廖礼强[1] 

机构地区：[1]重庆医科大学附属第一医院心内科,重庆市400016 [2]重庆市急救医疗中心,重庆市400014

出　　处：《中国组织工程研究与临床康复》2010年第27期5018-5022,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:间充质干细胞具有改善心脏功能的潜力,间充质干细胞移植后向心脏归巢的机制尚不完全清楚。目的:观察心肌梗死大鼠体外培养的心肌组织对间充质干细胞迁移的作用及可能机制。方法:建立大鼠心肌梗死模型,培养大鼠心脏组织块,密度梯度离心及贴壁筛选法分离培养骨髓间充质干细胞,24只SD大鼠随机抽签法分为心肌梗死组、假手术组及正常对照组。假手术组只穿线不结扎,正常对照组不做任何处理。荧光染料DAPI标记间充质干细胞,利用transwell模型进行共培养,共培养48h。荧光镜下计数迁移细胞数,CD34/CD44免疫细胞化学染色进行迁移细胞定性,免疫荧光检测迁移细胞CXCR4的表达情况,心脏组织切片行免疫组织化学染色检测基质细胞衍生因子1的表达情况并进行平均吸光度分析。结果与结论:心肌梗死组单位视野迁移细胞数显著高于假手术组(P<0.05),正常对照组未见迁移细胞。免疫细胞化学染色显示迁移细胞CD44阳性而CD34阴性,符合间充质干细胞特点,其膜受体CXCR4阳性表达。心肌梗死组及假手术组心脏切片基质细胞衍生因子1阳性表达,正常对照组心肌组织不表达基质细胞衍生因子1。心肌梗死组心脏组织基质细胞衍生因子1平均吸光度显著高于其他组别(P<0.05)。提示心肌梗死后大鼠心脏组织能促进间充质干细胞迁移,这一效应的实现可能与基质细胞衍生因子1-CXCR4轴的作用有关。BACKGROUND：Mesenchymal stem cells（MSCs） have the potential to improve cardiac function,but the mechanisms of MSCs homing to heart post-implantation are not completely clear.OBJECTIVE：To investigate the role and possible mechanisms of cardiac tissue mass cultured ex vivo which isolated from myocardial infarction rats on MSC migration.METHODS：Rat models of myocardial infarction were established and cardiac tissue masses were cultured.MSCs were isolated from bone marrow by density gradient centrifugation and adherence screening method.A total of 24 Sprague Dawley rats were randomly assigned to myocardial infarction,sham operation and normal control groups.The rats in the sham operation group were only threaded without deligation.The rats in the normal control group were left intact.MSCs were labeled with DAPI.A transwell model was used to co-culture for 48 hours.The number of migrating cells was counted under a fluorescence microscope.Quantitative cell count was conducted for CD34/CD44 on MSCs that were tested using immunocytochemistry staining under a fluorescence microscope.Immunofluorescent staining was used to evaluate CXCR4 expression on MSCs.Immunohistochemistry staining was used to detect expression of stromal cell-derived factor-1 on cardiac sections and mean absorbance was analyzed.RESULTS AND CONCLUSION：Number of migrating MSCs in myocardial infarction group was significantly greater compared with sham operation group（P 0.05）.No migrating cells were detectable in the normal control group.Immunocytochemical staining results have shown that migrating cells were positive for CD44,but negative for CD34,which were consistent with MSC characteristics.The membrane receptor CXCR4 was positively expressed on the migrating cells.Stromal cell-derived factor-1-positive expression was observed on cardiac tissue sections both in myocardial infarction and sham operation groups,rather than in normal control group.Mean absorbance of stromal cell-derived factor-1 was significantly higher in the myocardial in

关 键 词：心肌梗死 心肌组织 间充质干细胞 迁移 大鼠 

分 类 号：R394.2[医药卫生—医学遗传学]