Development of Human Induced Pluripotent Stem Cell Derived Co-Cultures of Cardiomyocytes and Endothelial Cells

摘要

Cardiovascular disease is associated with myocardial infarction, arteriosclerosis, and cardiac arrhythmias, which lead to the segmental loss of functional beating cardiomyocytes. Numerous studies have investigated regenerative therapies involving stem cell transplantation using hydrogel scaffolds. However, these therapies have had limited success due to poor donor cell engraftment and low survival. One major contributing factor to the lack of success is the limited vascularization within the ischemic cardiac tissue and the scaffold. In this study we started to address these limitations by creating a novel co-culture of cardiomyocytes and endothelial cells derived from human induced pluripotent stem (iPS) cells using TGFβ1 as a secondary differentiation factor. TGFβ1 was added to an existing protocol that differentiates human iPS cells into cardiomyocytes to increase the endothelial cell population. We targeted CD 105 since it is a transmembrane receptor for TGFβ1, an important modulator of mesodermal differentiation, and is necessary for TGFβ1/ALK1 signaling which promotes the proliferation of endothelial cells. We determined the distribution of differentiated mesodermal cells using cell markers specific to progenitor (CD 105+) and mature (CD31+) endothelial cells along with cardiomyocytes (CTnT+) through fluorescence microscopy and flow cytometry analysis. We then assessed the differentiation of these iPSC-CPCs after seeding them into a hydrogel system of hyaluronic acid (HyA) to promote a 3D co-culture system.