The Study Of Different Principles And Treatments To Induce The Differentiation Of BMSCs To Cardiomyocyte-like Cells

Objective:To investigate the differences of promoting the differentiation ofBMSCs to cardiomyocyte-like cells among the Activating blood and dissolvingblood-stasis Therapy, the Qi-invigorating and Blood-activation Therapy, theQi-invigorating method.Method:Choose48SD rats to intragastric administration in groups, andpreparate drug serum through picking blood from abdominal aorta; use the50SD ratsto proceed the extraction, incubation and isolation purification of BMSCs;60SDneonatal rats to proceed the extraction, incubation and determination of myocardialcells, fetch their supernate by centrifugation to preparate conditioned medium in orderto mimic the rats’ myocardial microenvironment of extracorporeal. Use the preparateconditioned medium to induction CD105+-BMSCs, then randomly divided into4groups to join the drug-containing serum after3days, the Western Medicine addeddrugs directly to affect cells. Use MTT assay to choose the best compoundingconcentration of each treatment groups; draw the growth curve of CD105+-BMSCs ofeach treatment groups in1to7days; use FCM (flow cytometry) to test theproliferation case of CD105+-BMSCs of each treatment groups in1to5days; useimmunocytochemical method to test the differentiation of CD105+-BMSCs’expression of myocardium protein CTNT, GATA-4in1to5days.Results:(1) The CD105+-BMSCs of each treatment groups’ PopulationDoubling Time (PDT) is24h, the cells remain latent adaptation on the first and thesecond day; then they turn into the logarithmic phase on the third day, then theirproliferation reach the peak on the fourth day, the cells remain platform stage in5to7days; the cell number of the activating blood and removing stasis group in the secondday is apparently higher than the other treatment groups and the DMEM control groupon remains persistent state;(2) Except the1stday, the proliferation index (pi) of activa- ting blood and removing stasis group at every time point were all the maximum, thedivision generation was also apparently higher than other groups;(3)The normal grouphas the weakly positive expression of myocardium protein CTNT and GATA-4, andhad no obvious change with the passage of time; on the1stday, the activating bloodand removing stasis group, the supplementing Qi and activating blood circulationgroup, the supplementing Qi group, the G-CSF group and the normal saline group allhad the weakly positive expression of CTNT and GATA-4,they changed littlecomparing with DMEM control group, thus had no statistical significance (P>0.05);on day2,3,4and5, the expression quantity of CTNT and GATA-4in the activatingblood and removing stasis group, the supplementing Qi and activating bloodcirculation group, the supplementing Qi group increased obviously, comparing withthe DMEM control group has great significance (P<0.05); the activating blood andremoving stasis group is better than the supplementing Qi and activating bloodcirculation group, the supplementing Qi group, has the statistical significance(P<0.05); the difference of supplementing Qi group and the supplementing Qi&activating blood has no statistical significance (P>0.05).Conclusion:Under the condition of mimic myocardial microenvironment, themethod of promoting blood circulation and removing blood-stasis, the method of Ben-efiting Qi and Activating Blood, the strengthen qi therapy all have the effect of prom-oting the division of CD105+-BMSCs, and promoting the differentiation of BMSCs tocardiomyocyte-like cells, but the effect of the method of promoting blood circulationand removing blood-stasis is most obvious among the three therapies.