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Combinational Overexpression of Foxa3 and Hnf4a Enhance the Proliferation and Prolong the Functional Maintenance of Primary Hepatocytes

  • MOLECULAR BIOLOGY OF THE CELL
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Abstract

In an in vitro culture system, primary hepatocytes usually display a low proliferation capacity, accompanied with a decrease of viability and a loss of hepatocyte-specific functions. Previous studies have demonstrated that the combination introductions of certain hepatocyte-specific transcription factors are able to convert fibroblasts into functional hepatocyte-like cells. However, such combinational usage of transcription factors in primary hepatocytes culture has not yet sufficiently studied. The forkhead box protein A3 (FoxA3) and hepatocyte nuclear factor 4α (Hnf4α) are liver-enriched transcription factors that play vital roles in the differentiation, and maintenance of hepatocytes. Thus, we simultaneously overexpressed the two genes, Foxa3 and Hnf4a, in rat hepatocytes and observed that the combinational augmentation of these two transcription factors have enhanced the proliferation and stabilized the hepatocyte-specific functions of primary hepatocytes over a long-term culture period.

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Funding

This research work was supported by grants from the National Science Foundation of China (U1804186 and 81671619), Henan province Foundation (202300410307, 212102310893, 212102310611, 22A180009 and 22A320043), Xinxiang City Foundation (GG2020009).

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Authors and Affiliations

  1. Stem Cell and Biotherapy Engineering Research Center of Henan, Henan Joint International Research Laboratory of Stem Cell Medicine, Xinxiang Medical University, 453003, Xinxiang, China

    J. Y. Fan, G. Dama, Y. L. Liu & J. T. Lin

  2. College of Life Science and Technology, Xinxiang Medical University, 453003, Henan, Xinxiang, China

    J. Y. Fan, Y. L. Liu, W. Y. Guo & J. T. Lin

  3. Shandong Tianchuan Precision Medical Technology Co. Ltd., 253084, Dezhou, China

    J. Y. Fan

  4. Department of Community Health, Advanced Medical and Dental Institute, Universiti Sains Malaysia, 13200, Kepala Batas, Malaysia

    G. Dama

  5. College of Biomedical Engineering, Xinxiang Medical University, 453003, Henan, Xinxiang, China

    J. T. Lin

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  1. J. Y. Fan
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  2. G. Dama
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  3. Y. L. Liu
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Contributions

Jinyu Fan and Ganesh Dama contributed equally to this work. Conceptualization, Jinyu Fan and Juntang Lin; methodology, Jinyu Fan, Ganesh Dama; investigation, Jinyu Fan, Ganesh Dama and Weiyun Guo; formal analysis, Jinyu Fan, Ganesh Dama and Weiyun Guo; resources, Juntang Lin; data curation, Jinyu Fan; preparation and writing original draft, Jinyu Fan; writing-revision and editing, Ganesh Dama and Yanli Liu; supervision, Juntang Lin; project administration, Juntang Lin; funding acquisition, Juntang Lin. The authors have approved the final version of this manuscript.

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Correspondence to J. T. Lin.

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Conflict of interest. All authors declared no competing interests.

Statement on the welfare of animals. This study was approved by the ethical committee and Animal Care Committee (no. 030032) of Xinxiang Medical University. All animal experiments were performed according to the guidelines of The Ministry of Science and Technology of the People’s Republic of China [(2006)398].

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Fan, J.Y., Dama, G., Liu, Y.L. et al. Combinational Overexpression of Foxa3 and Hnf4a Enhance the Proliferation and Prolong the Functional Maintenance of Primary Hepatocytes. Mol Biol 57, 661–669 (2023). https://doi.org/10.1134/S0026893323040039

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