Construction Of Engineered Cardiac Tissue In Vitro Using Collagen Scaffold: A Experimental Study

Objectives: To construct engineered cardiac tissue using collagen sponge as scaffold based on cardiac myocytes as seed cells derived from newborn rats.Contents: Rat tail collagen was isolated and made into collagen sponge scaffold, crosslinked by heat dehydration. Neonatal rat cardiac myocyte was isolated using trypsin and collagenase. The appropriate concentration of BrDU in the culture medium for the inhibition of fibroblast was determined by cell counter. Engineered heart tissue was constructed.Methods: Collagen solution was lyophilized to form sponge scaffold. The scaffold was crosslinked by heat dehydration and observed by SEM. Neonatal rat cardiac myocyte was isolated using trypsin and collagenase solution. The number of isolated cell was counted by cell counter. The morphology and pulsation of the cardiomyocyte were investigated by microscopy. The proliferation curve of fibroblast under the inhibiton of BrDU was surveyed by cell counter. The microstructure of engineered cardiac tissue was examined using SEM and HE.Results: The crosslinked collagen sponge scaffold had sustained its porous structure, and the degradability rate was decreased. The cardiac myocyte isolated by digestion using trypsin and collagenase had more number and could beat longer than that isolated using single enzyme. The proliferation of fibroblast was efficiently inhibited when the concentration of BrDU was between 0.13 and 0.16mmol/L. The cardiac myocytes began beating spontaneous on the 3rd day of culture. Then they connected one another, and tended to beat synchronously. The engineered cardiac tissue can beat spontaneously .Conclusions: Engineered cardiac tissue can be constructed using collagen sponge as scaffold based on cardiac myocytes as seed cells derived from newborn rats. Engineered heart tissue was similar to natural cardiac tissue.