Ott H C, Bonaros N, Marksteiner R, Wolf D, Margreiter E, Schachner T, Laufer G, Hering S
Department of Cardiac Surgery, University of Innsbruck, Anichstrasse, A-6020 Innsbruck, Austria.
Eur J Cardiothorac Surg. 2004 Apr;25(4):627-34. doi: 10.1016/j.ejcts.2003.12.031.
To prove whether intramyocardial transplantation of combined skeletal myoblasts (SM) and mononuclear bone marrow stem cells is superior to the isolated transplantation of these cell types after myocardial infarction in rats.
In 67 male Fischer rats myocardial infarction was induced by direct ligature of the LAD. Seven days postinfarction baseline echocardiography and intramyocardial cell transplantation were performed. Via lateral thoracotomy 200 microl containing either 10(7) SMs or 10(7) bone marrow-derived mononuclear cells (BM-MNC) or a combination of 5x10(6) of both cell types (MB) were injected in 10-15 sites in and around the infarct zone. In controls (C) 200 microl of cell-free medium were injected in the same manner. Before injection both cell types were stained using a fluorescent cell linker kit (PKH, Sigma). In addition, SMs were transfected with green fluorescent protein. Nine weeks postinfarction follow-up echocardiography was performed and animals were sacrificed for further analysis.
At baseline echocardiography there was no difference in left ventricular ejection fraction (LVEF; C, SM, BM-MNC, MB: 60.1+/-3.2, 53.3+/-10.2, 53.1+/-8.7, 49+/-9.0%) and left ventricular end diastolic diameter (LVEDD; C, SM, BM-MNC, MB: 6.5+/-0.8, 5.17+/-0.8, 5.77+/-1.4, 6.25+/-0.8 mm) between the different therapeutic groups. Eight weeks after cell transplantation LVEDD was significantly increased in all animals except those that received a combination of myoblasts and bone marrow stem cells (MB; C, SM, BM-MNC, MB: 7.7+/-0.6 mm, P=0.001; 7.7+/-1.5 mm, P<0.001; 7.7+/-1.1 mm, P=0.005; 6.6+/-1.7 mm, P=0.397. At the same time LVEF decreased significantly in the control group (C), stayed unchanged in animals that received bone marrow stem cells (BM-MNC) and increased in animals that received myoblasts (SM) and a combination of both cell types (MB; C, SM, BM-MNC, MB: 45.3+/-7.0%, P=0.05; 63.9+/-15.4%, P=0.044; 54.3+/-6.3%, P=0.607; 63.0+/-11.5%, P=0.039).
The present data show that the concept of combining SMs with bone marrow-derived stem cells may be of clinical relevance by merging the beneficial effects of each cell line and potentially reducing the required cell quantity. Further studies are required to identify the exact mechanisms underlying this synergy and to allow full exploitation of its therapeutic potential.
证实大鼠心肌梗死后，联合骨骼肌成肌细胞（SM）与单核骨髓干细胞进行心肌内移植是否优于单独移植这些细胞类型。
对67只雄性Fischer大鼠通过直接结扎左前降支诱导心肌梗死。梗死7天后进行基线超声心动图检查及心肌内细胞移植。经左胸切口，在梗死区域及其周边10 - 15个部位注射200微升含有10⁷个SM或10⁷个骨髓来源的单核细胞（BM - MNC）或两种细胞各5×10⁶个的组合（MB）。对照组（C）以同样方式注射2百微升无细胞培养基。注射前，两种细胞类型均使用荧光细胞连接试剂盒（PKH，Sigma）进行染色。此外，SM用绿色荧光蛋白转染。梗死9周后进行随访超声心动图检查，并处死动物进行进一步分析。
在基线超声心动图检查时，不同治疗组之间的左心室射血分数（LVEF；C、SM、BM - MNC、MB：60.1±3.2%、53.3±10.2%、53.1±8.7%、49±9.0%）和左心室舒张末期内径（LVEDD；C、SM、BM - MNC、MB：6.5±0.8毫米、5.17±0.8毫米、5.77±1.4毫米、6.25±0.8毫米）无差异。细胞移植8周后，除接受成肌细胞和骨髓干细胞组合（MB）的动物外，所有动物的LVEDD均显著增加（C、SM、BM - MNC、MB：7.7±0.6毫米，P = 0.001；7.7±1.5毫米，P < 0.001；7.7±1.1毫米；P = 0.005；6.6±1.7毫米，P = 0.397）。同时，对照组（C）的LVEF显著降低，接受骨髓干细胞（BM - MNC）的动物LVEF保持不变，接受成肌细胞（SM）和两种细胞组合（MB）的动物LVEF增加（C、SM、BM - MNC、MB：45.3±7.0%，P = 0.05；63.9±15.4%，P = 0.044；j54.3±6.3%，P = 0.607；63.0±11.5%，P = 0.039）。
目前数据表明，将SM与骨髓来源的干细胞联合的理念可能具有临床意义，通过融合各细胞系的有益作用并可能减少所需细胞数量。需要进一步研究以确定这种协同作用的确切机制，并充分发挥其治疗潜力。
