Mesenchymal stem cells culture12/30/2023 Cell sheet technology eliminates the problem of retention, as all MSCs remain at the implantation site. Suspensions of MSCs injected into the ischemic area of the heart often present underwhelming results, because of cell loss and a low retention of transplanted cells. High colonization rates and long survival times after transplanting are two key points in the curative effects of MSCs. Several clinical studies have described how the use of MSCs improved cardiac function and the regeneration of heart tissue in damaged hearts. MSCs secrete growth factors and chemokines which induce cardio-protection, stimulate angiogenesis, and reorganize the extracellular matrix in ischemic scar areas. MSCs have been used to treat damaged hearts as they offer a low immunogenic property, paracrine effect, and immunosuppression. It is well-known that heart tissue is difficult to repair itself, and adult cardiac cells tend not to divide, so it may be possible to repair ischemic areas by transplanting mesenchymal stem cells (MSCs). Recently, stem cell therapy has been developed and extensively investigated as an alternative therapeutic strategy for heart failure. Other surgical interventions, such as the implanting of mechanical ventricular assist devices, have high associated medical costs and can easily cause bleeding, infection, and other complications. However, the shortage of donor hearts limits the application of this procedure, which also requires lifelong immunosuppression. Heart transplant is the standard curative therapy for end-stage heart failure. Heart failure is the leading cause of death worldwide, despite improvements in management through mechanical and surgical therapeutic strategies. Since UC-MSCs have few human leukocyte antigen-II and major histocompatibility complex class I molecules, and are easy to isolate and culture, UC-MSC sheets have been proposed as a candidate for clinical applications to ischemic heart disease. At present, many types of MSCs have been considered as multipotent cells in the treatment of heart failure, such as bone marrow-derived mesenchymal stem cells (BM-MSCs), adipose-derived mesenchymal stem cells (AD-MSCs), umbilical cord-derived mesenchymal stem cells (UC-MSCs), and skeletal myoblasts (SMs). In this review, we summarized the research and the applications of MSC sheets to the treatment of ischemic heart tissue. The long-term retention of MSCs in the affected area was realized and significantly improved the curative effect. In combination with cell sheet technology, MSCs could be more easily transplanted to the ischemic area. A purified population of MSCs can be obtained 3 weeks after the initiation of culture.In recent years, mesenchymal stem cells (MSCs) have been used to improve cardiac function and attenuate adverse ventricular remodeling of the ischemic myocardium through paracrine effects and immunoregulation functions. When primary cultures become almost confluent, the culture is treated with 0.5 ml of 0.25% trypsin containing 0.02% ethylenediaminetetraacetic acid for 2 min at room temperature (25 degrees C). Nonadherent cells are removed carefully after 3 h and fresh medium is replaced. Mouse mesenchymal stem cells are generally isolated from an aspirate of BM harvested from the tibia and femoral marrow compartments, then cultured in a medium with Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) for 3 h in a 37 degrees C-5% CO(2) incubator. Our protocol is mainly on the basis of the frequent medium change in primary culture and diminishing the trypsinization time. We explain a protocol for straightforward isolation and culture of mesenchymal stem cells (MSCs) from mouse bone marrow (BM) to supply researchers with a method that can be applied in cell biology and tissue engineering with minimal requirements.
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