Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Ekaterini D Zacharis
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Fabian Bachinger
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Annabelle Wurmser
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Daniel Yamamoto
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Sandra Petrus-Reurer
Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Carola M Morell
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Dominika Dziedzicka
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Brandon T Wesley
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom
Imbisaat Geti
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Charis-Patricia Segeritz
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Miguel C de Brito
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Mariya Chhatriwala
Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Daniel Ortmann
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Kourosh Saeb-Parsy
Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom
Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge and NIHR Cambridge Biomedical Research Centre, Cambridge, United Kingdom; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom
Production of large quantities of hepatocytes remains a major challenge for a number of clinical applications in the biomedical field. Directed differentiation of human pluripotent stem cells (hPSCs) into hepatocyte-like cells (HLCs) provides an advantageous solution and a number of protocols have been developed for this purpose. However, these methods usually follow different steps of liver development in vitro, which is time consuming and requires complex culture conditions. In addition, HLCs lack the full repertoire of functionalities characterising primary hepatocytes. Here, we explore the interest of forward programming to generate hepatocytes from hPSCs and to bypass these limitations. This approach relies on the overexpression of three hepatocyte nuclear factors (HNF1A, HNF6, and FOXA3) in combination with different nuclear receptors expressed in the adult liver using the OPTi-OX platform. Forward programming allows for the rapid production of hepatocytes (FoP-Heps) with functional characteristics using a simplified process. We also uncovered that the overexpression of nuclear receptors such as RORc can enhance specific functionalities of FoP-Heps thereby validating its role in lipid/glucose metabolism. Together, our results show that forward programming could offer a versatile alternative to direct differentiation for generating hepatocytes in vitro.
