Journal: Scientific Reports

Article Title: Evidence for bystander signalling between human trophoblast cells and human embryonic stem cells

doi: 10.1038/srep11694

Figure Lengend Snippet: ( a ) Protocol for indirect exposure of target cells (fibroblasts or hES cells) to Co and Cr ions. ( b ) Protocol for direct exposure of target cells (fibroblasts or hES cells) to Co and Cr ions. ( c ) Analysis of DNA damage in the BeWo bi-layer and fibroblast cells using the alkaline comet assay. Three hundred cells (three repetitions of 100) were analysed at random per parameter per experiment. All experiments were repeated three times giving a total of 900 cells scored per parameter. ( d ) Measurement of total Co and Cr concentration in conditioned medium using inductively coupled plasma mass spectrometry (ICP-MS). Conditioned medium was collected from below nine BeWo barriers and from nine placenta explants (taken from three placentae). The data was compared by one-way ANOVA. When a P value of >0.05 was found post hoc Dunnett’s tests were used to compare each treatment group to the negative control. *P > 0.05, **P > 0.01, ***P > 0.001 when compared to the negative control. Centre values represent means. Error bars represent SEM.

Article Snippet: Primary human BJ skin fibroblasts (LGC Promochem UK) were maintained in T-75 flasks (Corning) in Minimal Essential Medium (Sigma-Aldrich) supplemented with 10% (v/v) foetal bovine serum (Gibco), 1% (v/v) antibiotic antimitotic solution (containing 1000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL, Sigma Aldrich), 0.1 mg/mL sodium pyruvate, 0.02M HEPES buffer and 2 mM L-glutamine (all from Sigma-Aldrich) at 5% CO 2 and 37 °C.

Techniques: Alkaline Single Cell Gel Electrophoresis, Concentration Assay, Clinical Proteomics, Mass Spectrometry, Negative Control