Preparation Of BM-MSC Multilayered Cell Sheet With Favorable Stem Cell Activity For Myocardial Infarction In A 3D Dynamic System

Purpose:The present study aims to construct a multilayered bone marrow-mesenchymal stem cell?BM-MSC?cell sheet by using a 3D dynamic culture system,and a rat MI model would be used to study the therapeutic effect and relative mechanisms of cell sheet.Method:The phospholipase A₂ decellularized method was used to prepare decellularized porcine pericardium?DPP?,and aspartic acid?Asp?was used to crosslink VEGF on DPP.The 3D multilayered structure was constructed with RAD16-I hydrogel,and a dynamic culture cell sheet?DCcs?or a static culture cell sheet?SCcs?was acquired by using a dynamic perfusion culture system or static culture condition for 48 h.Cell number,proliferation,apoptosis,surface markers expression,and growth factor levels of BM-MSC within cell sheets were examined.SCcs and DCcs were transplanted in a rat MI model,respectively,the cardiac function and morphology of infarcted myocardium were evaluated within 4 weeks.The survival of BM-MSC was traced,and the angiogenesis of infarcted myocardium was also evaluated.Result:The heterogeneous cells were efficiently removed from DPP,and the natural collagen were well-preserved.The DPP-Asp-VEGF scaffold was successfully prepared and VEGF stably released in 7 days.After a 48 h culture,DCcs exhibited higher cell density,better proliferation,superior paracrine abilities,and lower cell apoptosis compared to the SCcs.In vivo,compared to the MI control group,DCcs significantly reduced infarcted area and improved cardiac function.And a higher BM-MSC survival rate and better angiogenesis were observed in DCcs rather than SCcs group.Conclusion:A multilayered BM-MSC cell sheet with favorable stem cell bioactivity was successfully constructed by using a 3D dynamic culture system,and transplanting DCcs in a rat MI model significantly reduced the infarcted area,improved myocardial function,and facilitating angiogenesis of the infarcted myocardium.