| Objective: Attentive observation of intramyocardial injection of bone marrow mesenchymal stem cells (mesenchymal stem cells, MSCs) and endothelial progenitor cells (endothelial progenitor cells, EPCs) transplantation on cardiac function in rats with acute myocardial infarction, myocardial cell transcription factor gene, myocardial cell markers protein content, heart infarct capillary density effects.Comparison of combined transplantation MSCs and EPCs whether higher than that alone one of the cell transplantation, MSCs, and EPCs two kinds of cell transplantation whether there are differences between the results.Methods: 1. Take 2 weeks of clean grade healthy SD rats , extracted bone marrow , by density centrifugation extracted mononuclear cells, differential adhesion method and the different medium induction to obtain MSC and EPC. Flow cytometry screening, so that MSCs and EPCs can be purified. With DAPI (final concentration of 50mg / L) marked MSCs and EPCs. Observed two kinds of cell morphology. 2. Through the left anterior descending coronary artery ligation in rat myocardial infarction model building. 3. Groups of laboratory animals and cell transplantation: The model of myocardial infarction have been made,SD rats were randomly grouped for cell transplantation. Group A: MSCs transplantation group 1 week after the completion of myocardial infarction model , intramyocardial injection of MSCs suspension 200ul (5x106) have been marked through the outer membrane of myocardial cells; Group B: EPCs transplantation group 1 week after the completion of myocardial infarction model , intramyocardial injection of MSCs suspension 200ul (5x106) have been marked through the outer membrane of myocardial cells; group C: co-transplantation group 1 week after the completion of myocardial infarction model , intramyocardial injection of MSCs and EPCs suspension 200ul (5x106) have been marked through the outer membrane of myocardial cells; group D : control group 1 week after the completion of myocardial infarction model , intramyocardial injection of Medium DMEM suspension 200ul through the outer membrane of myocardial cells.4. 4 weeks after cell transplantation measured cardiac hemodynamics, Then rats were killed to take the heart of specimens for paraffin sections, fluorescence microscopy observation of DAPI labeled MSCs and EPCs, and counting, WithⅧfactor immunohistochemistry capillary density, measured by immunohistochemistry of cardiac troponin I and connexin 43, extracted myocardial tissue RNA, measuring cardiac transcription factor genes. 5. Statistical analysis: All experimental data with x±s said that the use of SPSS10.0 software processing the data. Multiple mean comparison with the single-factor analysis of variance. If they meet the homogeneity of variance, and further using Bonferroni pairwise comparison test, If the variance is incomplete, and using Tamhane's T2 pairwise comparison method. P f<0.05 for significant difference.Results: 1. Through differential adherent culture method to be more pure isolated mesenchymal stem cells and endothelial progenitor cells. 2. group A, group B and group C can see the DAPI labeled cells, compared with the control group at the same time, gross vascular density increases, left ventricular end-diastolic pressure (LVEDP) decreased ,left ventricular systolic pressure (LVSP), Maximum left ventricular pressure rate of rise and fall (±dp / dtmax), cardiac-specific transcription factor GATA-4, Nkx-2.5 and the VEGF gene semi-quantitative and cardiac troponin I and connexin 43 were significantly higher (P "0.05). 3. Cell transplantation compared between groups, the Joint transplantation group index change is more obvious; mesenchymal stem cell transplantation group when compared with the progenitor cells, endothelial progenitor cells in groups with higher levels of VEGF, capillary density increased more apparent; The mesenchymal transcription factors and of cardiac troponin levels higher than the progenitor transplantation group (P <0.05).Conclusion: The DAPI marks can be applied to stem cell transplantation. Cell transplantation group improvement of cardiac function, capillary rise, myocardial cell marker increased. combined transplantation is better compared with single-cell transplantation... |
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