BACKGROUND AND PURPOSE: Teratogenic substances induce adverse effects during the development of the embryo. Multilineage differentiation of human embryonic stem cells (hESCs) mimics the development of the embryo in vitro. Here, we propose a transcriptomic approach in hESCs for monitoring specific toxic effects of compounds as an alternative to traditional time-consuming and cost-intensive in vivo tests requiring large numbers of animals. This study was undertaken to explore the adverse effects of cytosine arabinoside (Ara-C) on randomly differentiated hESCs. EXPERIMENTAL APPROACH: Human embryonic stem cells were used to investigate the effects of a developmental toxicant Ara-C. Sublethal concentrations of Ara-C were given for two time points, day 7 and day 14 during the differentiation. Gene expression was assessed with microarrays to determine the dysregulated transcripts in presence of Ara-C. KEY RESULTS: Randomly differentiated hESCs were able to generate the multilineage markers. The low concentration of Ara-C (1 nM) induced the ectoderm and inhibited the mesoderm at day 14. The induction of ectodermal markers such as MAP2, TUBB III, PAX6, TH and NESTIN was observed with an inhibition of mesodermal markers such as HAND2, PITX2, GATA5, MYL4, TNNT2, COL1A1 and COL1A2. In addition, no induction of apoptosis was observed. Gene ontology revealed unique dysregulated biological process related to neuronal differentiation and mesoderm development. Pathway analysis showed the axon guidance pathway to be dysregulated. CONCLUSIONS AND IMPLICATIONS: Our results suggest that hESCs in combination with toxicogenomics offer a sensitive in vitro developmental toxicity model as an alternative to traditional animal experiments.
BACKGROUND AND PURPOSE: Teratogenic substances induce adverse effects during the development of the embryo. Multilineage differentiation of human embryonic stem cells (hESCs) mimics the development of the embryo in vitro. Here, we propose a transcriptomic approach in hESCs for monitoring specific toxic effects of compounds as an alternative to traditional time-consuming and cost-intensive in vivo tests requiring large numbers of animals. This study was undertaken to explore the adverse effects of cytosine arabinoside (Ara-C) on randomly differentiated hESCs. EXPERIMENTAL APPROACH: Human embryonic stem cells were used to investigate the effects of a developmental toxicant Ara-C. Sublethal concentrations of Ara-C were given for two time points, day 7 and day 14 during the differentiation. Gene expression was assessed with microarrays to determine the dysregulated transcripts in presence of Ara-C. KEY RESULTS: Randomly differentiated hESCs were able to generate the multilineage markers. The low concentration of Ara-C (1 nM) induced the ectoderm and inhibited the mesoderm at day 14. The induction of ectodermal markers such as MAP2, TUBB III, PAX6, TH and NESTIN was observed with an inhibition of mesodermal markers such as HAND2, PITX2, GATA5, MYL4, TNNT2, COL1A1 and COL1A2. In addition, no induction of apoptosis was observed. Gene ontology revealed unique dysregulated biological process related to neuronal differentiation and mesoderm development. Pathway analysis showed the axon guidance pathway to be dysregulated. CONCLUSIONS AND IMPLICATIONS: Our results suggest that hESCs in combination with toxicogenomics offer a sensitive in vitro developmental toxicity model as an alternative to traditional animal experiments.
Authors: Sabrina C Desbordes; Dimitris G Placantonakis; Anthony Ciro; Nicholas D Socci; Gabsang Lee; Hakim Djaballah; Lorenz Studer Journal: Cell Stem Cell Date: 2008-06-05 Impact factor: 24.633
Authors: Chris T Dee; Caroline S Hirst; Yu-Huan Shih; Vineeta B Tripathi; Roger K Patient; Paul J Scotting Journal: Dev Biol Date: 2008-05-24 Impact factor: 3.582
Authors: T Komori; H Yagi; S Nomura; A Yamaguchi; K Sasaki; K Deguchi; Y Shimizu; R T Bronson; Y H Gao; M Inada; M Sato; R Okamoto; Y Kitamura; S Yoshiki; T Kishimoto Journal: Cell Date: 1997-05-30 Impact factor: 41.582
Authors: Douglas A Hosack; Glynn Dennis; Brad T Sherman; H Clifford Lane; Richard A Lempicki Journal: Genome Biol Date: 2003-09-11 Impact factor: 13.583
Authors: Aldert H Piersma; Nancy C Baker; George P Daston; Burkhard Flick; Michio Fujiwara; Thomas B Knudsen; Horst Spielmann; Noriyuki Suzuki; Katya Tsaioun; Hajime Kojima Journal: Curr Res Toxicol Date: 2022-05-13
Authors: Anne K Krug; Raivo Kolde; John A Gaspar; Eugen Rempel; Nina V Balmer; Kesavan Meganathan; Kinga Vojnits; Mathurin Baquié; Tanja Waldmann; Roberto Ensenat-Waser; Smita Jagtap; Richard M Evans; Stephanie Julien; Hedi Peterson; Dimitra Zagoura; Suzanne Kadereit; Daniel Gerhard; Isaia Sotiriadou; Michael Heke; Karthick Natarajan; Margit Henry; Johannes Winkler; Rosemarie Marchan; Luc Stoppini; Sieto Bosgra; Joost Westerhout; Miriam Verwei; Jaak Vilo; Andreas Kortenkamp; Jürgen Hescheler; Ludwig Hothorn; Susanne Bremer; Christoph van Thriel; Karl-Heinz Krause; Jan G Hengstler; Jörg Rahnenführer; Marcel Leist; Agapios Sachinidis Journal: Arch Toxicol Date: 2012-11-21 Impact factor: 5.153
Authors: Eugen Rempel; Lisa Hoelting; Tanja Waldmann; Nina V Balmer; Stefan Schildknecht; Marianna Grinberg; John Antony Das Gaspar; Vaibhav Shinde; Regina Stöber; Rosemarie Marchan; Christoph van Thriel; Julia Liebing; Johannes Meisig; Nils Blüthgen; Agapios Sachinidis; Jörg Rahnenführer; Jan G Hengstler; Marcel Leist Journal: Arch Toxicol Date: 2015-08-14 Impact factor: 5.153
Authors: Kesavan Meganathan; Smita Jagtap; Vilas Wagh; Johannes Winkler; John Antonydas Gaspar; Diana Hildebrand; Maria Trusch; Karola Lehmann; Jürgen Hescheler; Hartmut Schlüter; Agapios Sachinidis Journal: PLoS One Date: 2012-08-28 Impact factor: 3.240
Authors: K Meganathan; S Jagtap; S P Srinivasan; V Wagh; J Hescheler; J Hengstler; M Leist; A Sachinidis Journal: Cell Death Dis Date: 2015-05-07 Impact factor: 8.469
Authors: B Zimmer; G Pallocca; N Dreser; S Foerster; T Waldmann; J Westerhout; S Julien; K H Krause; C van Thriel; J G Hengstler; A Sachinidis; S Bosgra; M Leist Journal: Arch Toxicol Date: 2014-04-02 Impact factor: 5.153