Abstract
Direct reprogramming represents an easy technique to generate induced hepatocytes (iHeps) from somatic cells. However, current protocols are accompanied by several drawbacks as iHeps are heterogenous and lack fully mature phenotypes of primary hepatocytes. Here, we established a polycistronic expression system to induce the direct reprogramming of mouse embryonic fibroblasts towards hepatocytes. The resulting iHeps are homogenous and display key properties of primary hepatocytes, such as expression of hepatocyte markers, albumin secretion, and presence of liver transaminases. iHeps also possess the capacity to repopulate decellularized liver tissue and exhibit enhanced hepatic maturation. As such, we present a novel strategy to generate homogenous and functional iHeps for applications in tissue engineering and cell therapy.
Highlights
Orthotopic liver transplantation is the only cure for severe acute and chronic liver diseases
We report the efficient generation of induced hepatocytes (iHeps) from mouse embryonic fibroblasts (MEFs) using a polycistronic system expressing transcription factors including Forkhead Box A3 (Foxa3), HNF1 Homeobox A (Hnf1α), and GATA Binding Protein 4 (Gata4)
RNA was isolated from MEFs, primary hepatocytes, and two dimension‐cultured iHeps using the RNeasy Micro Kit according to the manufacturers instructions (Qiagen, Hilden, Germany)
Summary
Orthotopic liver transplantation is the only cure for severe acute and chronic liver diseases. Direct reprogramming has been shown to allow the generation of induced hepatocytes (iHeps) from many types of somatic cells (Du et al, 2014; Huang et al, 2011; Sekiya & Suzuki, 2011) These iHeps acquired hepatocyte function to some extent and could extend the survival of mouse models with lethal liver disease after cell transplantation. We and others have reported that decellularized liver tissue could provide an excellent environment for the in vitro differentiation of hepatic stem cells (Vyas et al, 2018; Wang et al, 2011) as well as maintenance of primary hepatocytes (Soto‐Gutierrez et al, 2011).
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
