Undifferentiated murine embryonic stem cells used to model the effects of the blue–green algal toxin cylindrospermopsin on preimplantation embryonic cell proliferation

• Published in: Toxicon (Oxford) Vol. 106; pp. 79 - 88
• Main Authors: Reid, Katherine J., Lang, Kenneth, Froscio, Suzanne, Humpage, Andrew J., Young, Fiona M.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.11.2015
• Subjects: Algae; Animals; Bacterial Toxins - toxicity; Biological Assay - methods; Biomedical materials; Cell Differentiation - drug effects; Cell Proliferation - drug effects; Cercopithecus aethiops; COS Cells; Crystal violet cytotoxicity assay; Crystals; Cyanobacteria - chemistry; Cylindrospermopsin; Drinking water; Embryonic development; Embryonic Development - drug effects; Embryonic Stem Cells; In vivo testing; In vivo tests; Mice; Mouse embryonic stem cell; Neural differentiation; Reproducibility of Results; Surgical implants; Toxins; Uracil - analogs & derivatives; Uracil - toxicity; Crystal violet cytotoxicity assay; Embryonic development; Neural differentiation; Mouse embryonic stem cell; Cylindrospermopsin
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/j.toxicon.2015.09.022

Undifferentiated mouse embryonic stem cell (mES) proliferation in vitro resembles aspects of in vivo pre-implantation embryonic development. mES were used to assess the embryo-toxicity of cylindrospermopsin (CYN), a water contaminant with an Australian Drinking Water Guideline (ADWG) of 1 μg/L. mES exposed to 0–1 μg/mL CYN for 24–168 h were subjected to an optimised crystal violet viability assay. mES exposed to retinoic acid ± 1 μg/L CYN differentiated into neural-like cells confirmed by morphological examination and RT-PCR for Oct4, Brachyury and Nestin. The CYN No Observed Effect Concentration (OEC) was 0.5 μg/mL, the Lowest OEC was 1 μg/mL (p < 0.001, n = 3), and the IC50 was 0.86 μg/mL after 24 h. The ADWG 1 μg/L CYN did not affect differentiation or proliferation after 72 h, but decreased proliferation after 168 h (p < 0.05). We conclude that higher algal bloom-associated CYN concentrations have the potential to impair in vivo pre-implantation development, and the mES crystal violet assay has broad application to screening environmental toxins. •72 h exposure to drinking water CYN guideline 1 μg/L did not affect mES proliferation.•1 μg/L CYN did not affect mES differentiation.•High algal bloom CYN toxin levels have potential for preimplantation toxicity.•Australian Drinking Water CYN guideline 1 μg/L likely to be ‘safe’ for early embryos.•Embryonic stem cell assay has broad application to screening environmental toxins.