title: Evaluation of human embryonic stem cells and their differentiated fibroblastic progenies as cellular models for in vitro genotoxicity screening.
abstract: This study evaluated human embryonic stem cells (hESC) and their differentiated fibroblastic progenies as cellular models for genotoxicity screening. The DNA damage response of hESCs and their differentiated fibroblastic progenies were compared to a fibroblastic cell line (HEPM, CRL1486) and primary cultures of peripheral blood lymphocytes (PBL), upon exposure to Mitomycin C, gamma irradiation and H2O2. It was demonstrated that hESC-derived fibroblastic progenies (H1F) displayed significantly higher chromosomal aberrations, micronuclei formation and double strand break (DSB) formation, as compared to undifferentiated hESC upon exposure to genotoxic stress. Nevertheless, H1F cell types displayed comparable sensitivities to genotoxic challenge as HEPM and PBL, both of which are representative of somatic cell types commonly used for genotoxicity screening. Subsequently, transcriptomic and pathways analysis identified differential expression of critical genes involved in cell death and DNA damage response upon exposure to gamma irradiation. The results thus demonstrate that hESC-derived fibroblastic progenies are as sensitive as commonly-used somatic cell types for genotoxicity screening. Moreover, hESCs have additional advantages, such as their genetic normality compared to immortalized cell lines, as well as their amenability to scale-up for producing large, standardized quantities of cells for genotoxicity screening on an industrial scale, something which can never be achieved with primary cell cultures.
authors.affiliation: Stem Cell Laboratory, Faculty of Dentistry, National University of Singapore, Singapore; Department of Oral & Maxillofacial Surgery, National Dental Centre, SingHealth, Singapore. Electronic address: vinoth.kumar@ndcs.com.sg.
authors.forename: Kumar Jayaseelan
authors.initials: KJ
authors.lastname: Vinoth
authors.affiliation: Center of Excellence in Genomic Medicine Research, King Abdulaziz University, PO Box 80216, Jeddah 21589, Kingdom of Saudi Arabia; School of Anatomy, Physiology and Human Biology, The University of Western Australia, 35 Stirling Highway, Crawley WA 6009, Australia.
authors.forename: Jayapal
authors.initials: J
authors.lastname: Manikandan
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore; School of Bio Sciences & Technology, VIT University, Vellore 632104, India.
authors.forename: Swaminathan
authors.initials: S
authors.lastname: Sethu
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Lakshmidevi
authors.initials: L
authors.lastname: Balakrishnan
authors.affiliation: Department of Biological Sciences, Faculty of Science & Technology, Sunway University, No. 5, Jalan Universiti, Bandar Sunway, 47500 Selangor Darul Ehsan, Malaysia; Department of Biosystems Science & Engineering, ETH-Zurich, Mattenstrasse 26, Basel 4058, Switzerland.
authors.forename: Alexis
authors.initials: A
authors.lastname: Heng
authors.affiliation: Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Japan.
authors.forename: Kai
authors.initials: K
authors.lastname: Lu
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore; Tembusu College, University Town, National University of Singapore, Singapore. Electronic address: phsmph@nus.edu.sg.
authors.forename: Manoor Prakash
authors.initials: MP
authors.lastname: Hande
authors.affiliation: Stem Cell Laboratory, Faculty of Dentistry, National University of Singapore, Singapore. Electronic address: dencaot@nus.edu.sg.
authors.forename: Tong
authors.initials: T
authors.lastname: Cao
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects* / Q000528:radiation effects; D002460:Cell Line / Q000187:drug effects / Q000528:radiation effects; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000528:radiation effects; D005347:Fibroblasts / Q000187:drug effects* / Q000528:radiation effects; D005720:Gamma Rays; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D066298:In Vitro Techniques; D016685:Mitomycin / Q000633:toxicity; D009152:Mutagenicity Tests
chemical_list: D016685:Mitomycin; D006861:Hydrogen Peroxide
keywords: DNA; Differentiation; Microarray; Mitomycin C; Oxidative; Radiation; Repair; Stress