BMP2in Infarcted Myocardium Lysate Induced Bone Marrow Mesenchymal Stem Cells Induced Differentiation To Cardiomyocytes

ObjectiveDiscussion on bone morphogenetic protein-2and myocardial infarctionlysate simulation of the biological micro-environment differentiation intocardiomyocytes possibility; and bone morphogenetic protein-2in themicro-environmen in action。MethodsTake health clean level2weeks age of SD rats bone marrow mesenchymalstem cells (BMSCs)to culture in vitro, while establishing the modle of myocardialinfarction and prepared into infarcted myocardium lysate. The cultivation of thethird generation of BMSCs divided into five groups,which density and activitywas the same in, pure DMEM training group (A), BMP-2group (B), infarctedmyocardium lysate group (C), infarcted myocardium lysate+noggin-blockergroup(D), BMP2+infarcted myocardium lysate group (E), these five groupswere3days for the liquid,3weeks cell harvesting.Induced cells morphologicalobservation under the inverted phase contrast microscope; Usingimmunohistochemical techniques, to detect the induced cells a-sarcomeric actinand Cardiac specific troponin(cTnT); and the induced cells were analyzed bystatistics.ResultsUnder an inverted microscope BMSCs after induction of cell growth andmorphology observation: group A after induction of BMSCs cell morphology did not produce any significant change, always a fibroblast-like morphology. Bgroup1weeks after induction of BMSCs cell extended pseudopodia, cellorientation is consistent;3weeks after induction of cell-cell interactionconnection increases, although are less intensive, but has obvious directivity,partially visible sheet-like muscle filament-like structures, but no formsarcomere-like structure. C group1weeks after induction of BMSCs graduallyincreased in size in a long fusiform;3weeks after induction, visible are widelydistributed, but the direction is not consistent, aligned to less intensive smallpiece of muscle filament-like structures, and no form of sarcomere structure.Group D after induction of BMSCs form always were fibroblast-like, does nothave obvious change. E group1weeks after induction of BMSCs volumeincreased, mostly were morphologically consistent, aligned parallel to the longfusiform;3weeks after induction of cells were morphologically consistent longspindle, visible extensive patchy muscle filament-like structures, are arranged indense.Immunohistochemical identification (1)identification of BMSCs:grow well inthe second generation of bone marrow mesenchymal stem cells byImmunohistochemical staining for detection of surface antigen CD44, CD166positive, the positive rate of above97%.(2)BMSCs induced by3weeks aftercTnT, a-sarcomeric actin detection:induction3weeks, group A and D cardiacspecific protein staining was negative; B, C, E three group of cTnT, a-sarcomericaction positive expression and no significant difference among the three groups.(P>0.05).ConclusionsBMP-2, infarcted myocardium lysate can induce differentiation of bonemarrow mesenchymal stem cells into cardiomyocyte like cells; BMP-2ininfarcted myocardium lysate plays a role; BMP-2infarcted myocardium lysateinduced by simple compared to the efficiency of induced no significant difference.