脐血间质干细胞经肌源性诱导后细胞内游离钙的变化  

Change of Intracellular Free Calcium in Mesenchymal Stem Cells Derived from Umbilical Cord Blood after Myogenic Induction

作　　者：沈赛娥[1] 梅举[2] 丁芳宝[2] 钟竑[2] 鲍春荣[2] 马南[2] 

机构地区：[1]上海交通大学医学院附属新华医院麻醉科,上海200092 [2]上海交通大学医学院附属新华医院胸心外科,上海200092

出　　处：《中国胸心血管外科临床杂志》2011年第2期130-135,共6页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

摘　　要：目的研究3种不同的肌源性诱导方法对脐血间质干细胞（mesenchymal stem cells,MSCs）进行诱导后细胞内Ca2＋调控系统分化的影响。方法 2007年1月至2010年4月上海交通大学医学院附属新华医院采用5-氮杂胞嘧啶核苷（5-azacytidine,5-aza）、心肌裂解液和心肌诱导液分别诱导犬脐血MSCs。采用激光共聚焦显微镜检测5-aza、心肌裂解液和心肌诱导液诱导的细胞、心肌细胞和MSCs内Ca2＋荧光探针Fluo-3/AM结合的Ca2＋,每份细胞选取2~5个视野,每一种细胞观察记录30个视野,取每1 min摄得的10张图象的平均荧光强度值,以光密度值（OD）反映细胞内游离Ca2＋浓度;并连续监测细胞内游离Ca2＋浓度的变化。结果经3种方法诱导后细胞内游离Ca2＋浓度均高于心肌细胞内游离Ca2＋浓度（F=59.400,P=0.000）。5-aza、心肌裂解液和心肌诱导液诱导的细胞内游离Ca2＋浓度差异有统计学意义（F=18.988,P=0.000）。5-aza诱导的细胞内游离Ca2＋浓度与心肌裂解液诱导的细胞比较差异无统计学意义（OD 1 076.88±44.65 vs.1 040.90±37.48,P=0.186）,但其浓度高于心肌诱导液诱导的细胞（OD 1 076.88±44.65 vs.973.91±46.49,P=0.001）,心肌裂解液诱导的细胞内游离Ca2＋浓度高于心肌诱导液诱导的细胞（OD 1 040.90±37.48 vs.973.91±46.49,P=0.001）。3种诱导方法诱导后的细胞内游离Ca2＋呈自发性波动,与心肌细胞相似,但其频率和幅度有明显差异。5-aza、心肌裂解液诱导后的MSCs在KCl刺激下能使细胞内Ca2＋瞬时释放,心肌诱导液诱导后的MSCs对KCl也有反应,但不能诱发Ca2＋释放,而是使Ca2＋释放时间延长。结论 5-aza诱导、心肌裂解液和心肌诱导液诱导对MSCs的Ca2＋调控系统有分化作用,3种方法的效果不同,其产生差异的原因和效果的优劣需进一步实验阐明。Objective To study the influence of three different ways of myogenic induction on Ca2＋ regulation of mesenchymal stem cells（MSCs） derived from umbilical cord blood.Methods From January 2007 to April 2010,three different ways of myogenic induction including the adoptions of 5-azacytidine,extraction of myocardium,and myocardial differentiation medium were used to induce MSCs derived from the umbilical cord blood of dogs in Xinhua Hospital of Shanghai Jiaotong University.Confocal laser scanning microscope was used to detect cells induced by the three above-mentioned methods,cardiomyocytes and Ca2＋ combined with Fluo-3/AM inside the MSCs.For each group of cells,2 to 5 visual fields were chosen,and 30 visual fields were recorded for each kind of cells.The mean fluorescence intensity of ten images shot in one minute was used to reflect the concentration of free intracellular Ca2＋.Furthermore,the change of the concentration was continuously monitored by optical density（OD） value.Results After induction,the Ca2＋ concentration inside the MSCs was significantly higher than that inside the cardiomyocytes（F=59.400,P=0.000）.There was a statistical difference among the intracellular Ca2＋ concentration induced respectively by 5-azacytidine, extraction of myocardium,and myocardial differentiation medium（F=18.988,P=0.000）.No significant difference existed between the intracellular Ca2＋ concentration induced by 5-azacytidine and extraction of myocardium（OD value： 1 076.88±44.65 vs.1 040.90±37.48,P=0.186）,while the intracellular Ca2＋ concentration induced by 5-azacytidine was significantly higher than that induced by myocardial differentiation medium（OD value： 1 076.88±44.65 vs.973.91±46.49,P=0.001）,and the intracellular Ca2＋ concentration induced by extraction of myocardium was significantly higher than that induced by myocardial differentiation medium（OD value： 1 040.90±37.48 vs.973.91±46.49,P=0.001）.The concentration of intracellular Ca2＋ induced by all the three differen

关 键 词：肌源性诱导 脐血 间质干细胞 钙离子 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]