| Objective:1.To explore the method for isolation and cultivation of adipose tissue-derived mesenchymal stem cells(ADMSCs).2.To confirm that ADMSCs can survive,differentiate into cardiomyocyte-like cells in vivo,and improve heart function.3. To investigate the possible mechanisms of adipose tissue-derived mesenchymal stem cells(ADMSCs) transplantation on the angiogenesis in myocardial infarction rabbits.Methods:1.Isolating,culturing and labelling ADMSCs.The adipose tissue was o-btained from the epiloon of rabbits under the aseptic condition.ADMSCs were isolate-d by the method of digesting with typeâ… collagenase and adereing to the culture flas-k,and continuely cultured in vitro. The third passage of ADMSCs were used to ident-ify surface molecular marks. The third passage were labelled by DAPI,and then obse-rved by fluoresecent microscope.2.Treatment after acute myocardial infarction with ADMSCs transplantation.30 rabbits are randomly divided into three groups:shame operation with PBS injection(sham group,n=10),heart infarcted model with PBS inje- ction(control group.n=10),heart infarcted model with ADMSCs group (ADMSCs group,n=10). The third passage of ADMSCs labelled by DAPI were injected into the center and the border of the MI area after AMI 1h.The same volume of PBS was inje-cted into the MI area of the control group and sham group. Echocardiography was pe-rformed to evaluate the cardiac function before operation and 4 weeks after MI.After 4 weeks,the hearts were harvested.The presence of labelled cells was confirmed by fl-uorescent microscope,and differentiation of the labelled cells were observed by immu-nohistochemical stain of c TnI.The infarcted area were sectioned for HE stain and cap-illary were sectioned for immunohistochemical stain of CD34. The expression of V-EGF and bFGF in ischemic region were detected with RT-PCR.Results:1.Rabbits adipose tissue contained a great quantity of mesenchymal stem cells,which were quite easy to isolate and culture.24h after inoculation,many bigger cells had been adheret to the tisse culture flask.After 48h,the cells began to proliferate,after 5-7 d-ay, cells reached 80%confluence. After the passage,ADMSCs appeared to adopt a m-ore uniform fibroblast-like shape with directionality and regularity, similar to bone mar-row stromal cells.The round or oval cells decreased with the exchanging the media and passage. CD71nd CD44 were all positive in these cells,but CD34 and CD45 were negative. The marked rate of the third passage of ADMSCs labelled by DAPI was almost 100%under fluor-escent microscope.2. Compared with AMI control group,4 weeks after implantation ADMSCs group resulted in shorter LVIDd,LVIDs, and higher FS,EF(p<0.05).The la-belled cells were visible in the recipient heart,and some of them differentiated into ca-rdiac muscle(p<0.05).3. The microvessal density in the infarction regions treated with ADMSCs was higher than that in AMI group.The expression of bFGF and VEGF in t-he myocardial tissue of ADMSCs group was higher than that in AMI group and cont-rol group(p<0.05).Conclusion:1.In our study,an economic,simply and efficient way to isolate, cult-ure and labelling of ADMSCs from rabbit adipose tissue is established.2. Transplanta-tion allograftic ADMSCs can survive and differentiate into cardiomyocyte,promote angiogenesis in the infarcted or ischemic area of myocardium,and improve the heart function after MI.3. The transplantation of ADMSCs can promote the revasculariza- tion in infarcted regions, and improve the heart function by enhancing VEGF and bFGF synthesis.
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