title: Efficient Differentiation of Human Induced Pluripotent Stem Cell (hiPSC) Derived Hepatocyte-Like Cells on hMSCs Feeder.
abstract: BACKGROUND
  The use of stem cells is considered as an appropriate source in cell therapy and tissue engineering. Differentiation of human induced Pluripotent Stem Cells (hiPSCs) to Hepatocyte-like Cells (HLCs) on mouse embryonic fibroblasts (MEFs) feeders is confronted with several problems that hinder the clinical applications of these differentiated cells for the treatment of liver injuries. Safe appropriate cells for stem cell-based therapies could create new hopes for liver diseases. This work focused on the determination of a capacity/efficiency for the differentiation of the hiPSCs into Hepatocyte-like Cells on a novel human adult bone marrow mesenchymal stem cells (hMSCs) feeder.
  
  
  METHODS
  Undifferentiated human iPSCs were cultured on mitotically inactivated human adult bone marrow mesenchymal stem cells. A three-step differentiation process has been performed in presence of activin A which added for 3 days to induce a definitive endoderm formation. In the second step, medium was exchanged for six days. Subsequently, cells were treated with oncostatin M plus dexamethasone for 9 days to generate hepatic cells. Endodermic and liver-specific genes were assessed via quantitative reverse transcription-polymerase chain reaction and RT-PCR, moreover, immunocytochemical staining for liver proteins including albumin and alpha-fetoprotein. In addition, functional tests for glycogen storage, oil red examination, urea production and alpha-fetoprotein synthesis, as well as, cells differentiated with a hepatocyte-like morphology was also performed.
  
  
  RESULTS
  Our results show that inactivated human adult bone marrow mesenchymal stem cell feeders could support the efficient differentiation of hiPSCs into HLCs. This process induced differentiation of iPSCs into definitive endocrine cells that expressed sox17, foxa2 and expression of the specific genes profiles in hepatic-like cells. In addition, immunocytochemical analysis confirmed albumin and alpha-fetoprotein protein expression, as well as, the hiPSCs-derived Hepatocyte-like Cells on human feeder exhibited a typical morphology.
  
  
  CONCLUSIONS
  we suggested a successful and efficient culture for differentiation and maturation of hepatocytes on an alternative human feeders; this is an important step to generate safe and functional hepatocytes that is vital for regenerative medicine and transplantation on the cell-based therapies.
authors.affiliation: Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran ; Students'Sciences Research Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Naser
authors.initials: N
authors.lastname: Mobarra
authors.affiliation: Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Kouhkan
authors.affiliation: Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Hesari
authors.affiliation: Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Reyhaneh
authors.initials: R
authors.lastname: Lahmy
authors.affiliation: Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Mossahebi-Mohammadi
authors.affiliation: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran ; Microbial Biotechnology Laboratory, Department of Microbiology, School of Biology, College of Science, Tehran University, Tehran, Iran.
authors.forename: Ehsan
authors.initials: E
authors.lastname: Arefian
authors.affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Jaafarpour
authors.affiliation: Hematology-Oncology and Stem cell Transplantation Research Center, Shariati Hospital, Tehran university of Medical Science, Tehran,Iran.
authors.forename: Hajar
authors.initials: H
authors.lastname: Nasiri
authors.affiliation: Departments of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Pakzad
authors.affiliation: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Rezvan
authors.initials: R
authors.lastname: Tavakoli
authors.affiliation: Metabolic disorder Research center, Endocrinology and Metabolism, Molecular sciences institute, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Parvin
authors.initials: P
authors.lastname: Pasalar
keywords: Bone marrow mesenchymal stem cells; Hepatocyte-like Cells; Induced pluripotent stem cells