| AMI Mesenchymical stem cells are isolated culture,induced,labeled and transplanted into the infracted size.The purpose of the experiment is to study the influence of bone marrow mesenchymal stem cells(bMSCs) implantation into infarcted myocardium on blood vessel density and reducing apoptosis.METHOD The experiment is performed at the animal laboratory and the central laboratory,The First Affiliated Hospital of Anhui Mdical University between May 2006 and March 2008.①animal:totally 36 healthy mongrel canines are equally divided into control group and experimental group at random,18/each group.What to do with animals in the experiment is consistent with animal ethical standard.②bMSCs culture,inducing and labeling in vitro:10 ml bone marrow is aspirated from the posterosuperior iliac spine of canine with asepsis,then is isolated and purified by percoll density gradient centrifugation and then cultured in vitro by adhering to the culture plastic.The expression of cell surface antigen is detected via flow-cytometer. After being co-cultured with 5-azacytidine(5-Aza) for 24 hours,these cells are labeled by 5-bromodeoxyuridine(BrdU) and are to be applied.③bMSCs transplantation:the model of AMI is established in both groups.2 ml cell suspension of autologous bMSCs is implanted into four different positions in the acute myocardial site via topical injection around the infracted in experimental group while the same dose DMEM is injected into the correspond position in control group.④experiment evaluation:1,2 and 4 weeks after engraftation,the samples from experimental group and control group are collected to detect and analysis the cardiocyte apoptotic index(CAI) by TUNEL staining and blood vessel density by immunohistochemistry staining.RESULTS①The original generation cells and the passage cells culture in vitro:cells cultured in vitro adhere obviously after 24 h,cells show many kinds of shape such as round,fusiform,and irregularity;after passages,the form of cells is becoming to be conformity,being fusiform.The reproductive activity of bMSCs cultured in vitro is strong,bMSCs grow colony and the shape of cell colony is on "swirl",the growth curve of bMSCs is the shape of "S".②the identification of cell surface antigen:after passages of bMSCs,the positive rate of CD34 and CD31 on bMSCs cell surface is below 5%while the positive rate of CD44 and CD105 is above 90%.③the detection of cardiocyte apoptotic index:the sample slices derived from the transplantation sites are stained by BrdU immunofluorescence and the result is positive that prove that bMSCs transplanted survive in infarcted site.the cell nucleus of apoptotic cells are buffy by TUNEL staining,and compared with control group,cardiocyte apoptotic index (CAI) is much lower than that of control group(P＜0.01).④the matter of blood vessel density in infarcted site:blood vessel density in infarcted site in experiment group is much higher than those in control group(p＜0.05);and with the implantation time prolonging,it increases gradually,the distinction of tendency analysis has significance meaning.(F=439.588,p＜0.01).CONCLUSION:①bMSCs are isolated easily and have fairly proliferative ability,they grow fast when cultured in vitro,which will be an ideal seed cells for cell engraftment and cardial tissue engineering.②After implanted into the infarcted site,canin autologous bMSCs which survived can increase blood vessel density and reduce the numbers of cardiocytes apoptosis in damaged site.
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