title: Transferability of a modified embryonic stem cell test using a new endpoint for developmental neurotoxicity.
abstract: We developed and analyzed a new surrogate endpoint of the mouse embryonic stem cell test (EST) for developmental neurotoxicity. To determine the sensitivity, specificity, and transferability of the new endpoint, a pre-validation team from three independent laboratories optimized and standardized the protocol for neuronal differentiation of mouse embryonic stem cells (mESCs) by measuring the neuronal differentiation rates of mESCs under different culture conditions, such as the presence or absence of basic fibroblast growth factor (bFGF) in the growth media and varying lengths of culture. In addition, a component ratio of neuronal cells was measured by using flow cytometry analysis of β-III tubulin (Tuj1)-positive cells and real-time polymerase chain reaction analysis of microtubule-associated protein 2 (MAP2) mRNA. Our results showed that the best growth was achieved by culturing mESCs for 12 d in N2B27 medium without bFGF or ascorbic acid. Lead (II) acetate and aroclor 1254 were used to test the usefulness of the new endpoint. When we used the known ID(50) values for lead (II) acetate in the EST model, it was classified as non-embryotoxic; however, when we used the new ID(50) values that we determined in this study, it was classified as weakly embryotoxic. Aroclor 1254 and penicillin G were also classified as weakly embryotoxic and non-embryotoxic compounds, respectively, when cardiac and neuronal differentiation ID(50) values were used. Therefore, our new surrogate endpoint for developmental neurotoxicity is not only sensitive and specific but also transferable among laboratories.
authors.affiliation: National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration , Cheongwon-gun, Chungcheongbuk-do Republic of Korea.
authors.forename: Dae Hyun
authors.initials: DH
authors.lastname: Baek
authors.forename: Su-Yeon
authors.initials: SY
authors.lastname: An
authors.forename: Jae Hyun
authors.initials: JH
authors.lastname: Park
authors.forename: Youngju
authors.initials: Y
authors.lastname: Choi
authors.forename: Ki Dae
authors.initials: KD
authors.lastname: Park
authors.forename: Jin Wook
authors.initials: JW
authors.lastname: Kang
authors.forename: Kyoung Suk
authors.initials: KS
authors.lastname: Choi
authors.forename: Sung Hee
authors.initials: SH
authors.lastname: Park
authors.forename: Min Young
authors.initials: MY
authors.lastname: Whang
authors.forename: Jiyou
authors.initials: J
authors.lastname: Han
authors.forename: Jong-Hoon
authors.initials: JH
authors.lastname: Kim
authors.forename: Hyung Soo
authors.initials: HS
authors.lastname: Kim
authors.forename: Dongho
authors.initials: D
authors.lastname: Geum
authors.forename: Tae Moo
authors.initials: TM
authors.lastname: Yoo
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D020111:Chlorodiphenyl (54% Chlorine) / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000187:drug effects*; D007854:Lead / Q000633:toxicity; D051379:Mice; D041681:NIH 3T3 Cells; D009474:Neurons / Q000187:drug effects*
chemical_list: D020111:Chlorodiphenyl (54% Chlorine); D007854:Lead