摘要
目的:观察脐血间充质干细胞经5-氮杂胞苷诱导在体外定向分化为心肌样细胞的形态学及分子生物学特征。方法:选取2003-05/2005-04在郧阳医学院附属十堰市太和医院妇产科患者12例,脐血来源者家属及本人知情同意。取脐静脉血50mL,分离并培养骨髓间充质干细胞,用5-氮杂胞苷定向诱导向心肌样细胞分化。每日采用相差显微镜、透射电镜观察心肌样细胞形态学变化及超微结构特征,采用RT-PCR检测心房利钠肽、受磷蛋白、β-肌球蛋白重链表达。结果:①形态学变化:5-氮杂胞苷诱导后,约1周出现短棒状或珠状结构,2周排列具有方向性,3周有肌管样结构形成。②超微结构:诱导四五周细胞内含大量线粒体,可见心房颗粒、肌丝形成。③RT-PCR检测:经5-氮杂胞苷诱导四五周骨髓间充质干细胞有心房利钠肽、受磷蛋白、β-肌球蛋白重链表达。且心房利钠肽出现3条阳性带。结论:经5-氮杂胞苷诱导脐血间充质干细胞体外可分化为心肌样细胞,是细胞移植的优良供体。
AIM： To study morphological and molecular biological feature of differentiation of umbilical cord blood stem cells in vitro after induced by 5-asacytidine into cardiomyocytos. METHODS： A total of 12 patients were enrolled from Department of Obstetriea and Gynecology, Taihe Hospital of Shiyan City, Yunyang Medical College from May 2003 to April 2005. Family members and themselves of the cord blood source knew and agreed the fact. 50 mL umbilical venous blood was gained. Mesenchymal stem cells （MSCs） were isolated and cultured, and differentiated into cardiomyocytes after being identified by 5- azacytidine. Morphological change and ultrastructure feature of cardiomyocytes were obsoived with contrast phase microscope and transmission electron microscope daily. Expressions of atrial natriuretic polypeptide, phospholamhan and myosin heavy chain mRNA were examined with RTPCR. RESULTS： （1）Morphological change： The size of some cells had a stick or ball like one week after induced by 5-azacytidine. At week 2 the cells arranged directionally. At week 3 there was myotubes shape. （2）Ultrastructure： A mass of cytochondriome appeared in cells at weeks 4 and 5, and auricle granule and myoneme appeared. （3）RT-PCR examination： Expressions of atrial natriuretic polypeptide, phospholamban and β-myosin heavy chain mRNA in MSCs appeared at weeks 4 and 5＂ after induced by 5-asacytidine. Three positive strips appeared in atrial natriuretic polypeptide. CONCLUSION： After induced by 5-azacytidine, mesenchymal stem cells of cord blood that are good donators for cell transplant can differentiate into cardiomyocytes in vitro.
出处
《中国临床康复》
CSCD
北大核心
2006年第25期12-13,26,i0001,共4页
Chinese Journal of Clinical Rehabilitation
