| Object: To separate and purify MSCs and EPCs,to discuss the impact of MSCs co-transplantation with EPCs to the effect of myocardial ischemic rats.Methods:1.There were 50 adult male SD rats ,at first,five rats were selected as the source of donor bone marrow stem cells;40 rats by ligating the left anterior descending coronary artery were caused to establish myocardial ischemia model,the survival rats after making a successful model(total 30)were randomly divided into operation control group(n=6), MSCs group(n=8),EPCs group(n=8),MSCs and EPCs co-transplantation group(n=8),and other five were sham operation group(only open the chest and place thread without ligating). 2. MSCs and EPCs were purified and cultured in vitro。Took the third generation cells to transplant. 3. After 7～10 days of success of modeling, the rats in each group were transplanted with the MSCs and/or EPCs; control groups were injected with equivalent blank media. 4 weeks after cell transplantation,five groups rats'hemodynamic indexes were measured, heart slices were made to observe the myocardial tissue,the infarct size and the capllary density.Results: 1.40 rats died in the process of making three deaths;before and after transplantation each group rats had died;At last,when hemodynamic tests were taken, the operation group left 3 rats, MSCs group 6 rats, EPCs group 4 rats, Co-transplantation group 6 rats,and sham operation group had 5 rats.2.Use density gradient centrifugation and adherent screening to separate stem cells, and use DMEM and M199 culture medium to induced.3.After 4 weeks, the survival rats'hemodynamic indexes were that Co-transplantation group compared with operation group, MSCs group and EPCs group,the LVSP(127.3±9.5 vs 94.0±7.2 vs 112.7±10.1 vs 109.4±6.3)mmHg,+dp/dt(897.7±45.6 vs 440.5±61.8 vs 663.2±80.7 vs 540.4±58.5)mmHg/s and -dp/dt(610.6±59.7 vs 346.2±68.1 vs 547.0±60.7 vs 460.5±39.8)mmHg/s were obviously elevating (P <0.05); in contrast,the LVDP(6.3±0.6 vs 15.7±4.4 vs 9.7±1.8 vs 10.8±2.1)mmHg had decreased (P<0.01). There were no obvious difference between MSCs group and EPCs group (P>0.05).(3) The pathological examination revealed that sham operation group had no infarction; combination group's myocardial infarct size was the smallest (28.3±3.2)%,which was significantly lower than operation group (45.5±2.4)%( P≈0.001); It was also lower than MSCs group (34.0±6.1)% and EPCs group (36.5±3.3)%(all P<0.03); mealwhile, MSCs group and EPCs group were less than operation group (all P < 0.05), however, there was no significant difference between these two groups (P=0.10).The capillary density revealde that, combination group's figure (6.3±1.6)/200 times was the highest; EPCs group's (3.6±0.9) was higher than MSCs group's (2.1±0.3); and MSCs group's was also higher than operation group (1.1±0.3). The differences were evident between groups, all P<0.01.Conclusions: 1. Bone marrow stem cells'culture in vitro can be achieved in a large number of amplification, directed induction can take these cells into cardiomyocyte-like cells and endothelial cells. 2.MSCs and EPCs are mainly by improving hemodynamic index,reducing infarct size and promoting angiogenesis to improve cardiac function. 3. MSCs and EPCs co-transplantation can be used to treat ischemic heart disease, its consolidated effect is much better than these two cells alone.
|
PDF Full Text Request