title: Pancreatic islet differentiation of human embryonic stem cells by microRNA overexpression.
abstract: Development of stem cell-based therapies for the treatment of type 1 diabetes would provide a renewable supply of human β-cells. Human embryonic stem cells (ESCs) are considered to be one of the stem cell populations with sufficient proliferative capacity to achieve this goal. Currently, differentiation protocols directing ESCs toward a pancreatic fate employ a variety of expensive cytokines and inhibitors. With the known significance of microRNAs in islet development, we present a novel and cost-effective strategy in which miR-375 overexpression promotes pancreatic endocrine differentiation in hESCs in the absence of any extrinsic factors. miR-375 has been shown to be a key regulator of pancreatic development and function in zebrafish, mouse and human. In this study, hESCs were transduced with lentiviral vectors containing human miR-375 precursor and aggregated to form human embryoid bodies (hEBs) for up to 21 days. Morphological assessment, immunocytochemistry and DTZ staining confirmed that miR-375-induced hEBs have similar characteristics to those of mature islets. In addition, the dynamic expression profile of endodermal marker Foxa2 and endocrine-specific genes, including HNF4α, Pdx1, Pax6, Nkx6.1, Glut2 and insulin, were detected by quantitative real-time PCR. Finally, insulin release upon glucose stimulation was detected in our differentiated clusters. The data presented here demonstrate the feasibility of using microRNAs to direct differentiation into the pancreatic lineage. Copyright © 2013 John Wiley & Sons, Ltd.
authors.affiliation: Department of Genetics, Faculty of Biology Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Reyhaneh
authors.initials: R
authors.lastname: Lahmy
authors.affiliation: Department of Haematology, School of Medicine, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Mohammad H
authors.initials: MH
authors.lastname: Sanati
authors.affiliation: Department of Genetics, Faculty of Biology Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Behmanesh
authors.affiliation: Department of Genetics, Faculty of Biology Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Kouhkan
authors.affiliation: Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Naser
authors.initials: N
authors.lastname: Mobarra
mesh_terms: D000943:Antigens, Differentiation / Q000096:biosynthesis*; D002454:Cell Differentiation; D002460:Cell Line; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000943:Antigens, Differentiation; C540933:MIRN375 microRNA, human; D035683:MicroRNAs
keywords: diabetes; direct differentiation; genetic modification; human embryonic stem cell; miR-375; microRNA