The Study of Myocardial-like Tissue Engineering Constructed by Fibroblasts of Ischemia Myocardium

Abstract: Fibroblasts of ischemia myocardium (FIMs) were obtained from the ischemia myocardium of adult Wistar rats and induced by 5-azacytidine (5-aza) to be differentiated to cardiomyocyte-like cells (CLCs)， which were identified by indirect immunofluorescence against cardiac Troponin T (C-TnT) and α-sarcomeric actin (α- SA) antibodies and transmission electron microscope (TEM). Real-time PCR was performed to assay expressions of both angiotensinogen (AGT) and brain natriuretic peptide (BNP)mRNA in CLCs and FIMs. Scanning electron microscope (SEM) was used to evaluate two different scaffolds of acellular colon muscle film (ACMF) scaffold and Bio-Gide/Bio-Gide Perio (Bio-Gide) scaffold. CLCs were seeded on both ACMF scaffold and Bio-Gide scaffold and cultured for 5~7d. Indirect immunofluorescence against C-TnT and α-SA antibodies， SEM observation was done respectively to identify the myocardial-like tissue engineering. The results found that the cytoplasms of CLCs contained rich myofilaments， and were positive for C-TnT and α-SA. Real-time PCR revealed that the expressions of both AGT and BNP mRNA were much higher in CLCs than those in FIMs (P<0.05). CLCs showed elongated and/or polyhedral type， arranged in multilayers and stretched on ACMF scaffold and Bio-Gide scaffold to form myocardial-like stratums displayed rich myofilaments in the cytoplasms and were positive for C-TnT and α-SA. It was suggested that FIMs could differentiate into CLCs and be constructed myocardial-like tissue engineering on the different scaffolds.

CSTR: 32200.14.cjcb.2010.03.0006