| Objective : To separate, purificate and amplify swine bone arrow-derived mesenchymal stem cells (MSCs) through two methods that direct adherent and density gradient , then compared the cultivation efficiency of the two methods. To explore the feasibility and effectiveness of autologous transplanted MSCs joint bFGF after acute myocardial infarction. To provide a theoretical basis for clinical and experimental methods.Methods : took the swine bone marrow with the two separate methods that a simple direct adherent and density gradient centrifugation combined adherent to culture swine bone marrow mesenchymal stem cells and amplificate in vitro , then observed the shape of cell through the microscope and drawn its growth curve.Diannan swine 20, ligated the second diagonal branch of the left anterior descending thoracic to make the acute myocardial infarction model. All of these were divided into three groups randomly: group 1 with acute myocardial infarction (5). Determinated left ventricular ejection fraction (LVEF) by echocardiography after operation one day , determinated by selective coronary angiography after operation one week. and intracoronary infused non MSCs of saline injection by catheter, group 2 autologous transplantation of MSCs acute myocardial infarction (6) : Determinated left ventricular ejection fraction (LVEF) by echocardiography after operation one day , determinated by selective coronary angiography after operation one week. and intracoronary infused saline containing autologous MSCs by catheter. group 3 acute myocardial infarction autologous transplantation of MSCs joint pcDNA3 - bFGF (6) : Determinated left ventricular ejection fraction (LVEF) by echocardiography after operation one day , determinated by selective coronary angiography after operation one week. and intracoronary infused saline containing bFGF and MSCs by catheter. 4 weeks after administration, measured by echocardiography in left ventricular ejection fraction (LVEF)), and assessed left ventricular systolic function . oberserved coronary collateral of angiogenesis by selective coronary angiography again.Results : (1)the speed of collected the cell to proliferate is slow by direct adherent, and the speed of that is faster by density gradient centrifugation, and later all of these are spindle cells. (2) after ligated LAD , ECG .myocardium enzymology and selective coronary angiography all confirmed that the pig model of acute myocardial infarction is successful; (3) The left ventricular ejection fraction (LVEF) of the group that of MSCs and the group that of MSCs and pcDNA3-bFGF were higher than the control group by echocardiography four weeks after the operation.Conclusion : (1)the speed of culture MSCs by density gradient centrifugation combined adherent is faster than that of by a simple direct adherent.the MSCs that culture in vitro growed is stable, and the proliferation is stronge. (2) through imported cultured autologous MSCs by coronary, local micro-environment can be stimulated peripheral homing to myocardial infarction, its riched collateral circulation and improved heart function. The transplantation of MSCs joint bFGF to cure acute myocardial infarction may become a new treatment methods. |
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