Abstract 11: Xenogen-free In Vitro Differentiation and Expansion of Human Pluripotent Stem Cell-Derived Cardiovascular Progenitor Cells

Abstract

Background: Human pluripotent stem cell-derived cardiovascular progenitor cells (hPSC-CPCs) represent a tractable option for cell-based therapy for heart disease. However, to be clinically relevant, these cells must be derived under good manufacturing practices (GMP)-compatible conditions and produced in great enough quantities to treat adult patients. Here we sought to demonstrate for the first time the generation and expansion of clinically relevant numbers of hPSC-CPCs in xenogen-free protocol. Methods and Results: GMP-grade human induced pluripotent stem cells (GMP-hiPSCs) and human embryonic stem cells (H1 and H9) were dissociated into single cells and cultured in low attachment dishes to differentiate into CPCs in StemPro medium including small molecules and human cytokines with high efficiency of 86%, 80% and 66% for GMP-hiPSCs, H1 and H9, respectively (Figure 1). All hPSC-CPCs possessed trilineage differentiation potentials, as shown by differentiation into endothelial and smooth muscle cells and functional cardiomyocytes (Figure 2). Moreover, sorted hPSC-CPCs expanded >5 fold in 10 days in xenogen-free conditions while still maintaining trilineage differentiation potential and an efficiency of ~70% (Figure 3). Conclusions: Here we demonstrate a xenogeny-free CPC derivation and expansion protocol that can generate clinically relevant numbers of GMP-grade cardiovascular progenitors that could be used in a clinical setting.