Abstract
Direct conversion of cells through the overexpression of a set of transcription factors (TFs) is a promising approach for generating desired cell types from abundant and easily accessible cells. While previous studies have reported the generation of hepatic cells from fibroblasts, there is growing interest in understanding the mechanisms of hepatic reprogramming and its applications in regenerative medicine.
In this study, we demonstrated that overexpression of Yamanaka factors can convert mouse fibroblasts, in the presence of an optimized hepatic culture medium, into cells that closely resemble hepatocytes under in vitro conditions.
By examining the effects of 20 candidate transcription factors, we identified a combination of three factors—Hnf4a, Cebpa, and Nr1i2—that are capable of directing fibroblasts toward the hepatic lineage. When used alongside Yamanaka factors, these TFs significantly enhanced the efficiency of hepatic reprogramming.
The induced hepatocyte-like cells (iHep) generated using this method exhibited key hepatocyte-specific characteristics, including:
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Expression of liver-specific markers
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Glycogen storage
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Albumin secretion
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Urea production
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Low-density lipoprotein (LDL) uptake
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Indocyanine green (ICG) uptake
The production of iHep cells through these novel approaches can provide new insights into the molecular nature of hepatocyte differentiation and pave the way for the development of cell-based therapies for liver diseases.
Keywords:
Direct conversion; liver-specific transcription factors; hepatocyte-like cells; Yamanaka factors
