title: Evaluation of developmental toxicity using undifferentiated human embryonic stem cells.
abstract: An embryonic stem cell test (EST) has been developed to evaluate the embryotoxic potential of chemicals with an in vitro system. In the present study, novel methods to screen toxic chemicals during the developmental process were evaluated using undifferentiated human embryonic stem (hES) cells. By using surface marker antigens (SSEA-4, TRA-1-60 and TRA-1-81), we confirmed undifferentiated conditions of the used hES cells by immunocytochemistry. We assessed the developmental toxicity of embryotoxic chemicals, 5-fluorouracil, indomethacin and non-embryotoxic penicillin G in different concentrations for up to 7 days. While expressions of the surface markers were not significantly affected, the embryotoxic chemicals influenced their response to pluripotent ES cell markers, such as OCT-4, NANOG, endothelin receptor type B (EDNRB), secreted frizzled related protein 2 (SFRP2), teratocarcinoma-derived growth factor 1 (TDGF1), and phosphatase and tensin homolog (PTEN). Most of the pluripotent ES cell markers were down-regulated in a dose-dependent manner after treatment with embryotoxic chemicals. After treatment with 5-fluorouracil, indomethacin and penicillin G, we observed a remarkable convergence in the degree of up-regulation of development, cell cycle and apoptosis-related genes by gene expression profiles using an Affymetrix GeneChips. Taken together, these results suggest that embryotoxic chemicals have cytotoxic effects, and modulate the expression of ES cell markers as well as development-, cell cycle- and apoptosis-related genes that have pivotal roles in undifferentiated hES cells. Therefore, we suggest that hES cells may be useful for testing the toxic effects of chemicals that could impact the embryonic developmental stage.
authors.affiliation: Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, 361-763, Republic of Korea.
authors.forename: Eui-Man
authors.initials: EM
authors.lastname: Jung
authors.forename: Yeo-ul
authors.initials: YU
authors.lastname: Choi
authors.forename: Hong-Seok
authors.initials: HS
authors.lastname: Kang
authors.forename: Hyun
authors.initials: H
authors.lastname: Yang
authors.forename: Eui-Ju
authors.initials: EJ
authors.lastname: Hong
authors.forename: Beum-Soo
authors.initials: BS
authors.lastname: An
authors.forename: Jun-young
authors.initials: JY
authors.lastname: Yang
authors.forename: Ki Hwan
authors.initials: KH
authors.lastname: Choi
authors.forename: Eui-Bae
authors.initials: EB
authors.lastname: Jeung
mesh_terms: D000954:Antigens, Surface / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000187:drug effects*; D005472:Fluorouracil / Q000633:toxicity; D020869:Gene Expression Profiling; D006801:Humans; D007213:Indomethacin / Q000633:toxicity; D010400:Penicillin G / Q000633:toxicity; D060888:Real-Time Polymerase Chain Reaction; D018675:Toxicity Tests / Q000379:methods
chemical_list: D000954:Antigens, Surface; D010400:Penicillin G; D005472:Fluorouracil; D007213:Indomethacin
keywords: cell toxicity; developmental toxicant; differentiation; human embryonic stem cells; microarray