Comparative assessment of toxic responses in 3D embryoid body differentiation model and mouse early embryos treated with
- PDF / 10,115,986 Bytes
- 17 Pages / 595.276 x 790.866 pts Page_size
- 74 Downloads / 152 Views
REPRODUCTIVE TOXICOLOGY
Comparative assessment of toxic responses in 3D embryoid body differentiation model and mouse early embryos treated with 5‑hydroxytryptophan Olga Gordeeva1 · Andrey Gordeev2,3 Received: 25 July 2020 / Accepted: 10 September 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Pluripotent stem cells recapitulate in vitro the early developmental stages and are considered promising cell models for predictive developmental toxicity studies. To investigate the consistency between adverse drug effects on early development and the early stages of embryonic stem cell differentiation in three-dimensional (3D) in vitro culture, the toxic responses to 5-hydroxytryptophan (5-HTP; 0.5–2 mM) were evaluated in early mouse embryos and the embryoid body (EB) differentiation model. 3D architectures, developmental and differentiation dynamics and the cell death rates were analyzed in early mouse embryos (E2.5–E5.5) and EBs at 1 and 6 days of differentiation using a combination of confocal immunofluorescence microscopy with high content imaging analysis and quantitative gene expression analysis. Comparative analysis of toxic responses in early embryos and EBs revealed a similar dose- and stage-dependent decrease in the 5-HTP toxic effects during development and differentiation. The integral toxic responses in the early embryos and EBs were significantly dependent on their 3D architecture and cellular composition. Treatment with 5-HTP (1 mM and above) induced developmental arrest, growth inhibition, and increased cell death in the early embryos without the trophoblasts (E2.5) and those with impaired trophoblasts and in early EBs, whereas later embryos and EBs were more resistant due to the protection of the extraembryonic tissues. This study demonstrates that the EB differentiation model is a relevant 3D-model of early mammalian development and can be useful for the predictive evaluation of toxic and teratogenic effects in embryos at the preimplantation and early post-implantation developmental stages. Keywords 3D cell models · Developmental toxicity · Stem cells · Embryoid body · 5-HTP · Embryo
Introduction Pluripotent stem cells can differentiate into various somatic and germ cells and can recapitulate the morphogenetic events of early mammalian development in vitro. Therefore, pluripotent stem cells are considered unique and promising in vitro cell model of human and animal development for * Olga Gordeeva [email protected] 1
Laboratory of Cell and Molecular Mechanisms of Histogenesis, Kol’tsov Institute of Developmental Biology, Russian Academy of Sciences, 26 Vavilov Street, Moscow 119334, Russia
2
National Institutes of Health’s National Library of Medicine, Bethesda, MD 20852, USA
3
Medical Science and Computing, Bethesda, MD 20852, USA
predictive developmental toxicity and reproductive biology studies. A powerful technological platform based on pluripotent stem cells can be created for testing the effects of new drugs and environmental toxicants on all the c
Data Loading...
