The Study Of Derivation From Bone Marrow Stem Cells Of Adult Into A Cardiomyocyte Cells In Vitro

Objective:The sufferer's myocardial will be dead after AMI,Although recently research mean: After AMI,Few quantity themyocardial cell occurrence abruption and hyperplasia. But can'tthe complete repair myocardial organization. So the myocardialcell quantity reduces after AMI, the fiber scar tissuehyperplasy, the myocardial structure reconstruction, cause theheart failure. Currently the treatment means :thrombolysis andaeteria coronaria, implant frame Can only recover thereperfusion of the blood ,Prevent from the extension of themyocardium. But the different body heart transplantation hardclinic expansion because of donatorbody limit;graft rejectivereaction and High expenses. Method:A dult human bone marrow mesenchymal stem cells (MSCs) fromposterior superior iliac spine was gained on bone marrowaspiration. Separate the liquid separation to get single cellin the lymphoid cell of pecoll(1.073 g/ L) 。 Low-glucoseDulbecco's modified Eagle's medium (DMEM) ,Inoculate 10% humanserum of DMEM,37 ℃ s,5% CO2 hatches of incubator to becultivanted and changed the Liquid, passage.ALL cell will beobsered in contrast phase microscope. After three thassages, The cells are inoculate in sixborecultivante planks and diveded to tow groups,one groupinduced in 5-azacytidine 5*10mmol/L 5aza ( Sigma ) for 20hours.then hatched in the formal nutritive medium. The othergroups are hatched in the formal nutritive medium.The mediumwas changed twice a week thereafter until the experiment wasterminated 28days or 32days.All the cells were prepared for immunohistochemistry,transmission electron microscopy, RT-PCR.Results: The appearance Characteristics :The beginning inoculatedcells are as circular or oval types, regard circular asprinciple, the cells grow quickly after passage becomegradually long, looked like long spindle-shaped fibroblasticcells and began to form colonies. The induced cells growslowly, return small and become short,become graduallyuniform, to grow concentrately,There are afterbirth syrupsinside the endochylema,like the brush , ocontiguumfsbetween the cells.Immunohistochemistry :Tow group cells were positive forCD44,and in the traet grouop few cells were positive fora-action Trophonin T.Transmission electron microscopy: The cells' nucleus enlarge,tow nucleus are acquired ,there are a kind structurelikeintercalary disc among the cells, the bioblast bubblesand glycogen granules in the cytoplasm.there also a fewstructures like microfilament, but arrange not enoughintensive, the direction uniform is bad, expressing for theimmature muscle cells.RT-PCR:The mRNA expression of a-cardiac action , Cardiactropponin T. All values were normalized to Lyceraldehyde-3-phosphate dehydrogenase (GAPDH) Each measure of geneexpression in the experimental group was significantly higherthan its respective measure in the control group.Conclution:First step confirmation hMSC at the body was inducedproperly outside, can the centripetal muscle cell divide.Forclinical stem cell the application provides the experimentaccording to.Induced cells were immature cardiac muscle cells.the bioblast bubbles found in the Induced cells mayse ofthe toxicity of 5-aza.