[Acquirement and evaluation of XXXX ventricular extracellular matrix].

@entity19 @entity214
Cardiac extracellular matrix (ECM), generated from the process of decellularization, has been widely considered as an ideal source of biological scaffolds. However, current ECM preparations are generally difficult to be applied to generate cardiac tissue. Our research was aimed to improve decellularization protocols to prepare cardiac ECM slices. Adult @entity19 ventricular tissues were embedded in low melting agarose and cut into 300  m slices, and then were divided randomly into three groups: normal cardiac tissue, @entity214 treated group (0.1% @entity214 ) and @entity214 +Triton X-100 treated group (0.1% @entity214 +0.5% Triton X-100). Total RNA content and protein content quantification, HE staining and immunostaining were used to evaluate the removal of cell components and preservation of vital ECM components. Furthermore, @entity19 embryonic stem cell-derived cardiomyocytes (mES-CMs) and @entity19 embryonic fibroblasts (MEFs) were co-cultured with ECM slices to evaluate biocompatibility. The relative residual RNA and protein contents of ECM slices significantly decreased after decellularization. HE staining showed that @entity214 +Triton X-100 treatment better destroyed cellular structure and removed nuclei of ECM slices, compared with @entity214 treatment. Immunostaining showed that collagen IV and laminin were better preserved and presented better similarity to original cardiac tissue in ECM slices acquired by @entity214 +Triton X-100 treatment. However, collagen IV and laminin were significantly decreased and arranged disorderly in @entity214 treated group. We observed effective survival (>=  12 days) of MEFs and mES-CMs on ECM slices acquired by @entity214 +Triton X-100 treatment, and signs of integration, whereas those signs were not found in @entity214 treated group. We concluded that, compared with traditional @entity214 method, new combined protocol ( @entity214 +Triton X-100) generated ECM slices with better component and structural preservation, as well as better biocompatibility.
