Autospecies and Post–Myocardial Infarction Sera Enhance the Viability, Proliferation, and Maturation of 3D Cardiac Cell Culture

Abstract

The limited ability of cardiac muscle to regenerate after an extensive myocardial infarction (MI) and the scarcity of cardiac donors have fueled the field of cardiac tissue engineering as a potential therapeutic approach to enhance cardiac function in post-MI patients. We are exploring the ex vivo bioengineering of cardiac muscle tissue by seeding isolated cardiac cells within alginate scaffolds and supplementing the culture with "smart" media. The hypothesis investigated herein is that sera derived from autospecies and from post-MI animals contain agents that might induce cell proliferation, survival, and maturation in vitro. The results of the metabolic activity of the neonatal cardiac cell constructs (6.4-51x10(6) cells/cm(3)), as measured by MTT viability assay, indicated a significant advantage (p < 0.05) to the constructs supplemented with serum from normal and post-MI adult rats compared to fetal calf serum (FCS) supplementation. H&E staining and alpha-sarcomeric actin immunofluorescence staining revealed thick viable cardiac cell clusters (150-300 microm), with abundant 3D architecture in the cardiac cell constructs supplemented with post-MI and normal adult rat serum. The number of cells positively immunostained with Ki-67, a cell proliferation marker, was significantly higher in post-MI adult rat serum-supplemented cultures compared to negative results in the FCS-supplemented culture. The results presented in this study indicate that media supplemented with post-MI adult rat serum and normal adult rat serum compared to FCS have a significant advantage in the regeneration of injured cardiac tissue.