title: Undifferentiated murine embryonic stem cells used to model the effects of the blue-green algal toxin cylindrospermopsin on preimplantation embryonic cell proliferation.
abstract: 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.
authors.affiliation: Department of Medical Biotechnology, School of Medicine, Flinders University, Bedford Park, Adelaide, South Australia 5042, Australia; Australian Water Quality Centre, SA Water, 250 Victoria Square, Adelaide, South Australia 5001, Australia.
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Reid
authors.affiliation: Department of Medical Biotechnology, School of Medicine, Flinders University, Bedford Park, Adelaide, South Australia 5042, Australia; South Australian Department of Health, Government of South Australia, Adelaide 5000, Australia.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Lang
authors.affiliation: South Australian Department of Health, Government of South Australia, Adelaide 5000, Australia.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Froscio
authors.affiliation: Australian Water Quality Centre, SA Water, 250 Victoria Square, Adelaide, South Australia 5001, Australia.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Humpage
authors.affiliation: Department of Medical Biotechnology, School of Medicine, Flinders University, Bedford Park, Adelaide, South Australia 5042, Australia; Flinders Fertility, Flinders Medical Centre, Bedford Park, Adelaide, South Australia 5042, Australia. Electronic address: Fiona.Young@flinders.edu.au.
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Young
mesh_terms: D000470:Alkaloids; D000818:Animals; D001427:Bacterial Toxins / Q000633:toxicity*; D001681:Biological Assay / Q000379:methods; D019556:COS Cells; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002522:Chlorocebus aethiops; D000458:Cyanobacteria / Q000737:chemistry; D000087522:Cyanobacteria Toxins; D047108:Embryonic Development / Q000187:drug effects*; D053595:Embryonic Stem Cells; D051379:Mice; D015203:Reproducibility of Results; D014498:Uracil / Q000031:analogs & derivatives* / Q000633:toxicity
chemical_list: D000470:Alkaloids; D001427:Bacterial Toxins; D000087522:Cyanobacteria Toxins; C089595:cylindrospermopsin; D014498:Uracil
keywords: Crystal violet cytotoxicity assay; Cylindrospermopsin; Embryonic development; Mouse embryonic stem cell; Neural differentiation