Preparation And Preliminary Evaluation Of Decellularized Mouse Cardiac Textracellular Scaffold

Objective: The construction of three-dimensional microenvironment composites were made with BMSCs as seed cells and mice acellular heart scaffold as carrier.To evaluate the compose,component and internal structure of bone marrow mesenchymal stem cells(BMSCs)and acellular heart three-dimensional scaffold.To explore the biological characteristics of cell proliferation and differentiation of bone marrow mesenchymal stem cells in the three-dimensional microenvironment.To provide experimental evidence and the experimental basis of which carrying out related to tissue engineering and applied basic research.Methods: Optimize the three-dimensional microenvironment scaffolds materials: Preparation of acellular heart scaffold,trypsin as acellular reagent,SDS and Triton X-100 as detergent.The rat hearts were subjected to immersion shock in order to achieve cell removal rapidly.The acellular heart scaffold were observed under HE staining,Super differential photometer,scanning electron microscopic,Masson staining,immunohistochemical staining.Construction of cell scaffold composite by BMSCs in vitro,selected BMSCs by 7 days or 14 days co culture in the scaffold.The BMSCs heart scaffold complex were observed under HE staining,scanning electron microscope and real-time fluorescence quantitative PCR.Results: The heart scaffold looks like translucent cystic with trypsin,SDS and Triton X-100.No nucleus left with the HE staining.Acellular scaffold structure with a porous network and composed with fiber by SEM.To detect the protein component of acellular heart scaffold,the total DNA is 43.75�0.03ng/?L.The acellular scaffold main composed with blue reticular collagen fiber by Masson staining.Collagen type I with major part and collagen type III with a few part compose acellular scaffold,no expression of nuclear,myoglobin,actin with Immunohistochemical staining.The cells dispersed inside the scaffold and the cells number of 14 days co culture BMSCs scaffold were larger than 7 days by HE staining.BMSCs adhere in the surface of scaffold fiber tightly.The expression of ?-MHC and GATA-4 were higher with the induce of Myocardial tissue lysate in the BMSCs scaffold complex than in the BMSCs.Conclusion: In this study,the modified acellular method can effectively remove the cell components of mouse heart,and retain the extracellular collagen I type and III type.It means that in the condition of preserving the original spatial structure of the heart,the heart acellular scaffold has good biocompatibility,can maintain a low differentiated state and active proliferation.Well in simulating the in vivo tissue structure in a specific three-dimensional micro-environment culturing system.And can promote the directed differentiation of bone marrow mesenchymal stem cells.