Effects of qi supplementing, blood activating,and blood stasis resolving medicinals on differentiation of bone mesenchymal stem cells to cardiomyocyte-like cells in microenvironment of rat myocardium: a laboratory study in vitro*
2016, 39 (2):
Objective To investigate the effects of different therapeutic formulas on the differentiation of bone marrow mesenchymal stem cells (BMSC) to cardiomyocyte-like cells in rat myocardium of in vitro micro-environment. Methods 40 SD rats randomly divided into blood-activating stasis-resolving group, qi supplementing blood-activating group, qi supplementing group, and blank group were administered orally Chinese medicinals to obtain madicated serum; another 60 SD rats were used to extract, incubate, isolate and purify BMSC; SD newborn rats were used to extract and incubate myocardial cell, then to establish an external myocardial micro-environment. MTT assay was used to select the optimal compounding concentration of each treatment group. Flow cytometry was used to measure the proliferation of CD105+-BMSC of each treatment group. Immunocytochemical method was used to test the CD105+-BMSC’s expression of myocardium protein CTNT, GATA-4 while immunofluorescence technique was used to measure the expression of MHC. Results The doubling time of CD105+-BMSC of each treatment group was 24 h. The cells numbers in blood-activating and stasis-resolving group were significantly higher than those in control group and other treatment groups from the 2nd day, and remained steady. All the proliferation index of blood-activating and stasis-resolving group at every time point were the highest except the 1st day. The number of cell division was also significantly more than other groups: qi-supplementing and blood-activating group, qi-supplementing group, blood activating and stasis resolving group. From the 2nd day to 5th day, the expressions in the blood-activating stasis-resolving group, qi-supplementing blood-activating group, qi-supplementing group increased markedly with statistical significance compared with the blank group (P<0.05), blood-activating stasis-resolving group was superior to the other two groups(P<0.05). There were no statistical significance between the qi-supplementing blood-activating group and qi-supplementing group (P>0.05). Conclusion Under the external mimic myocardial micro-environment condition, all therapeutic formulas could promote CD105+-BMSC’s differentiation. They also could promote CD105+-BMSC’s metaplasia to the cardiomyocyte-like cells, however, the blood-activating stasis-resolving medicinal appears to be most effective.
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