Comparative induction of pluripotency in human umbilical vein endothelial cells and dermal fibroblasts and further differentiation into Hepatocytes

Abstract

Generating induced pluripotent stem cells (iPSCs) is a standardized technique. To date iPS cells have been generated from HUVECs using viral-based approaches. We employ an episomal plasmid-based approach to generate integration-free iPSCs (E-iPSCs) from human somatic cells. In this study, we compared the generation of E-iPSC lines from human umbilical vein endothelial cells (HUVEC) and fetal foreskin derived fibroblast (HFF1). The efficiency of inducing pluripotency in HFF1 was 0.03% compared to 2.5% in HUVEC. This implies that the efficiency of reprogramming HUVECs is 83-fold higher than in HFF1 cells. Additionally, the kinetics of reprogramming was much faster using HUVECs, i.e. three weeks for the stabilization of E-iPSC colonies compared to HFF1 cells which needed four months. The E-iPSCs from both somatic cell types were fully characterized and are comparable to human embryonic stem cells (hESCs). Both E-iPSC lines express pluripotency associated transcription factors as OCT4, NANOG, SOX2 and also the surface markers SSEA-4, TRA-l-60, TRA-1 – 81 and TRA-2 – 49 but not SSEA-1. Additionally, they have the ability to differentiate to all cell types representative of the three germ layers endoderm, ectoderm and mesoderm in vitro (by formation of embryoid bodies) and in vivo (teratoma formation in immunodeficient mice).