Grouping of histone deacetylase inhibitors and other toxicants disturbing neural crest migration by transcriptional profiling

Design of a high-throughput human neural crest cell migration assay to indicate potential developmental toxicants

“…Then toxicants were added and cells were allowed to migrate for 48 h Dreser et al, 2015). We maintained the 48-h toxicant exposure period in the new assay format, but we introduced a small modification: to avoid well-towell variations in the cell number at the start of the assay (due to 48 h of proliferation), we slightly modified the protocol by plating higher cell numbers (95,000 cells/cm 2 instead of 66,000 cells/ cm 2 ) and initiating toxicant exposure and migration one day after plating (Fig.…”

“…In a next step, two environmental toxicants known to inhibit NCC migration in the original MINC assay (Dreser et al, 2015) were tested. Both As 2 O 3 and CdCl 2 inhibited migration in the high nM range, but it was not entirely clear whether this effect could be distinguished from general cytotoxicity (Fig.…”

“…S3B). For this initial, rough evaluation, we adopted benchmark values that had proven useful in other studies (Zimmer et al, 2014;Dreser et al, 2015). For viability, a maximal variation of ≤10%, i.e., a reduction from 100% to 90% was deemed acceptable.…”

Design of a high-throughput human neural crest cell migration assay to indicate potential developmental toxicants_suppl

“…Several known developmental toxicants were positive in the original MINC assay. A screen of recently marketed drugs (Zimmer et al, 2014) and a further exploration of environmental toxicants (Dreser et al, 2015) identified additional NCC toxicants, such as polychlorinated biphenyls and interferon-beta.…”

“…Although a semi-automatic image quantification tool already has been established (Dreser et al, 2015), the throughput of the scratch assay is limited by manual image acquisition. Moreover, the assay is highly dependent on the experimenter's skills to achieve reproducible scratch properties.…”