title: Forced expression of Hnf4a induces hepatic gene activation through directed differentiation.
abstract: Embryonic stem (ES) cells are capable of unlimited self-renewal and have a diverse differentiation potential. These unique features make ES cells as an attractive source for developmental biology studies. Having the mature hepatocyte in the lab with functional activities is valuable in drug discovery studies. Overexpression of hepatocyte lineage-specific transcription factors (TFs) becomes a promising approach in pluripotent cell differentiation toward liver cells. Many studies generate transgenic ES cell lines to examine the effects of specific TFs overexpression in cell differentiation. In the present report, we have addressed whether a suspension or adherent model of differentiation is an appropriate way to study the role of Hnf4a overexpression. We generated ES cells that carried a doxycycline (Dox)-inducible Hnf4a using lentiviral vectors. The transduced cells were subjected to induced Hnf4a overexpression through both spontaneous and directed differentiation methods. Gene expression analysis showed substantially increased expression of hepatic gene markers, particularly Ttr and endogenous Hnf4a, in transduced cells differentiated by the directed approach. These results demonstrated that forced expression of TFs during directed differentiation would be an appropriate way to study relevant gene activation and the effects of overexpression in the context of hepatic differentiation.
authors.affiliation: Cellular and Molecular Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran.
authors.forename: Neda
authors.initials: N
authors.lastname: Yahoo
authors.affiliation: Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Electronic address: Pournasr@RoyanInstitute.org.
authors.forename: Behshad
authors.initials: B
authors.lastname: Pournasr
authors.affiliation: Department of Biological Sciences and Biotechnology, School of Science, University of Kurdistan, Sanandaj, Iran.
authors.forename: Jalal
authors.initials: J
authors.lastname: Rostamzadeh
authors.affiliation: Cellular and Molecular Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran. Electronic address: farfath@gmail.com.
authors.forename: Fardin
authors.initials: F
authors.lastname: Fathi
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051557:Hepatocyte Nuclear Factor 4 / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic
chemical_list: D051557:Hepatocyte Nuclear Factor 4; C496593:Hnf4a protein, mouse; D049452:Green Fluorescent Proteins
keywords: Directed differentiation; Embryonic stem cells; Hepatic genes; Hnf4a; Overexpression