Hepatocyte-like Cells Generated by Direct Reprogramming from Murine Somatic Cells Can Repopulate Decellularized Livers

Overview

Journal Biotechnol Bioeng

Publisher Wiley

Specialty Biochemistry

Date 2018 Jul 1

PMID 29959867

Citations 7

Authors

Chen Chen

Iris Pla-Palacin

Pedro M Baptista

Peng Shang

Loes A Oosterhoff

Monique E van Wolferen

Louis C Penning

Niels Geijsen

Bart Spee

Affiliations

Soon will be listed here.

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.

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References

Chen Y, Wong P, Sjeklocha L, Steer C, Sahin M . Mature hepatocytes exhibit unexpected plasticity by direct dedifferentiation into liver progenitor cells in culture. Hepatology. 2011; 55(2):563-74. PMC: 3268884. DOI: 10.1002/hep.24712. View

Baum C, Hegewisch-Becker S, ECKERT H, Stocking C, Ostertag W . Novel retroviral vectors for efficient expression of the multidrug resistance (mdr-1) gene in early hematopoietic cells. J Virol. 1995; 69(12):7541-7. PMC: 189692. DOI: 10.1128/JVI.69.12.7541-7547.1995. View

Jackson M, Taylor A, Jones E, Forrester L . The culture of mouse embryonic stem cells and formation of embryoid bodies. Methods Mol Biol. 2010; 633:1-18. DOI: 10.1007/978-1-59745-019-5_1. View

Takahashi K, Yamanaka S . Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006; 126(4):663-76. DOI: 10.1016/j.cell.2006.07.024. View

Kelaini S, Cochrane A, Margariti A . Direct reprogramming of adult cells: avoiding the pluripotent state. Stem Cells Cloning. 2014; 7:19-29. PMC: 3931695. DOI: 10.2147/SCCAA.S38006. View

Wang Y, Nicolas C, Chen H, Ross J, De Lorenzo S, Nyberg S . Recent Advances in Decellularization and Recellularization for Tissue-Engineered Liver Grafts. Cells Tissues Organs. 2017; 204(3-4):125-136. DOI: 10.1159/000479597. View

Kumar M, Keller B, Makalou N, Sutton R . Systematic determination of the packaging limit of lentiviral vectors. Hum Gene Ther. 2001; 12(15):1893-905. DOI: 10.1089/104303401753153947. View

Huang P, Zhang L, Gao Y, He Z, Yao D, Wu Z . Direct reprogramming of human fibroblasts to functional and expandable hepatocytes. Cell Stem Cell. 2014; 14(3):370-84. DOI: 10.1016/j.stem.2014.01.003. View

Schambach A, Bohne J, Baum C, Hermann F, Egerer L, von Laer D . Woodchuck hepatitis virus post-transcriptional regulatory element deleted from X protein and promoter sequences enhances retroviral vector titer and expression. Gene Ther. 2005; 13(7):641-5. DOI: 10.1038/sj.gt.3302698. View

10.

Nantasanti S, Spee B, Kruitwagen H, Chen C, Geijsen N, Oosterhoff L . Disease Modeling and Gene Therapy of Copper Storage Disease in Canine Hepatic Organoids. Stem Cell Reports. 2015; 5(5):895-907. PMC: 4649105. DOI: 10.1016/j.stemcr.2015.09.002. View

11.

Kim J, Lee S, Li L, Park H, Park J, Lee K . High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. PLoS One. 2011; 6(4):e18556. PMC: 3084703. DOI: 10.1371/journal.pone.0018556. View

12.

Vyas D, Baptista P, Brovold M, Moran E, Gaston B, Booth C . Self-assembled liver organoids recapitulate hepatobiliary organogenesis in vitro. Hepatology. 2017; 67(2):750-761. PMC: 5825235. DOI: 10.1002/hep.29483. View

13.

Gowda S, Desai P, Hull V, Math A, Vernekar S, Kulkarni S . A review on laboratory liver function tests. Pan Afr Med J. 2011; 3:17. PMC: 2984286. View

14.

Amand M, Hanover J, Shiloach J . A comparison of strategies for immortalizing mouse embryonic fibroblasts. J Biol Methods. 2019; 3(2):e41. PMC: 6706133. DOI: 10.14440/jbm.2016.110. View

15.

Cirillo L, Lin F, Cuesta I, Friedman D, Jarnik M, Zaret K . Opening of compacted chromatin by early developmental transcription factors HNF3 (FoxA) and GATA-4. Mol Cell. 2002; 9(2):279-89. DOI: 10.1016/s1097-2765(02)00459-8. View

16.

Cui J, Renaud H, Klaassen C . Ontogeny of novel cytochrome P450 gene isoforms during postnatal liver maturation in mice. Drug Metab Dispos. 2012; 40(6):1226-37. PMC: 3362787. DOI: 10.1124/dmd.111.042697. View

17.

LICHTSTEINER S, Schibler U . A glycosylated liver-specific transcription factor stimulates transcription of the albumin gene. Cell. 1989; 57(7):1179-87. DOI: 10.1016/0092-8674(89)90055-x. View

18.

Soto-Gutierrez A, Zhang L, Medberry C, Fukumitsu K, Faulk D, Jiang H . A whole-organ regenerative medicine approach for liver replacement. Tissue Eng Part C Methods. 2011; 17(6):677-86. PMC: 3103054. DOI: 10.1089/ten.TEC.2010.0698. View

19.

Huang P, He Z, Ji S, Sun H, Xiang D, Liu C . Induction of functional hepatocyte-like cells from mouse fibroblasts by defined factors. Nature. 2011; 475(7356):386-9. DOI: 10.1038/nature10116. View

20.

Sekiya S, Suzuki A . Direct conversion of mouse fibroblasts to hepatocyte-like cells by defined factors. Nature. 2011; 475(7356):390-3. DOI: 10.1038/nature10263. View