Partially Digested Adult Cardiac Extracellular Matrix Promotes Cardiomyocyte Proliferation In Vitro

Stimulating or maintaining the proliferative capacity of postnatal mammalian cardiomyocytes is a major challenge to cardiac regeneration. Previously, it is found that fetal cardiac extracellular matrix (ECM) can promote neonatal rat cardiomyocyte proliferation in vitro better than neonatal or adult ECM. It is hypothesized that partial digestion of adult ECM (PD-ECM) would liberate less crosslinked components that promote cardiomyocyte proliferation, similar to fetal ECM. Neonatal rat cardiac cells are seeded onto substrates coated with adult rat cardiac ECM that has been solubilized in pepsin-HCl for 1, 3, 6, 12, 24, or 48 h. Cardiomyocyte proliferation and fold-change in numbers from 1 to 5 d are highest on 1 and 3 h PD-ECM compared to other conditions. Sarcomeres tend to mature on 24 and 48 h PD-ECM where low proliferation is observed. 3 h PD-ECM is primarily composed of Fibrillin-1, Fibrinogen, and Laminins while 48 h PD-ECM is dominated by Collagen I. Our results suggest that adult ECM retains regenerative cues that may be masked by more abundant, mature ECM components. PD-ECM provides a simple yet powerful approach to promoting cardiomyocyte proliferation.